Template:Kyoto/Notebook/DMS

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Revision as of 21:14, 26 September 2014

7/31

Restriction Enzyme Digestion

Nakamura and Matsumoto

Sample/(µL)		EcoR1/(µL)	Xba1/(µL)	Spe1/(µL)	Pst1/(µL)	Buffer/(µL)		BSA/(µL)	MilliQ/(µL)	Total/(µL)
3/24 dddD PCR product	3.2	1	-	1	-	H	3	-	21.8	30
pSB1C3	18.5	1	-	1	-	H	3	-	6.5	30
pSB1C3 control	6.2	-	-	-	-	H	1	-	2.8	10

8/1

Gel Extraction

Murata and Yasuda

Lane	Sample
1	1kbp ladder
2	3/24 dddD PCR product E,S
3	3/24 dddD PCR product E,S
4	Blank
5	pSB1C3 E,S
6	pSB1C3 E,S
7	Blank
8	1kbp ladder
9	3/24 dddD PCR product control
10	pSB1C3 control

Sample	Concentration/(µg/mL)	260/280	260/230
PSB1C3	13.3	1.90	-
dddD	10.8	1.64	0.15

Ligation

Yasuda

Vector/(µL)		Insert/(µL)		Buffer/(µL)		Ligase/(µL)		MilliQ/(µL)	Total/(µL)
8/1 PSB1C3 gel extraction product 2047bp	1.5	8/1 dddD gel extraction product 2545bp	6.9	10xT4Ligase	2	T4Ligase	1	8.6	20

Transformation

Murata

Sample/(µL)		CompetentCells/(µL)		Total/(µL)	Medium
PSB1C3-dddD	3	コンピ名	30	33	SOC

8/2

PCR Cloning

Murata

dddD without Pre-Suf	2x KAPA HiFi Hotstart Ready Mix/(µL)	F Pre-dddD PCR product 62/(µL)	Rv Spe1-dddD PCR product 68/(µL)	MilliQ/(µL)	Total/(µL)
pick up	12.5	0.75	0.75	11	25

Predenature	Denature	Annealing	Extension
95°C	98°C	65°C	72°C
5min	20sec	15sec	50sec

Denature-Annealing-Extension 25cycle

DNA Purification

Murata

8/2 Pre-dddD-Suf PCR product

Colony PCR

Honda,Nakamura and Matsumoto

Master Mix

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
15	0.6	0.6	13.8	30

Predenature	Denature	Annealing	Extension
94°C	94°C	55°C	68°C
2min	30sec	30sec	24sec

Denature-Annealing-Extension 30cycle

Number	Sample
1	pSB1C3-dddD(1)
2	pSB1C3-dddD(2)
3	pSB1C3-dddD(3)

Concentration Measurement

Nakamura and Matsumoto

Sample	Concentration/(µg/mL)	260/280	260/230
dddD PCR product	245	1.59	2.17

Restriction Enzyme Digestion

Nakamura and Matsumoto

Sample/(µL)		EcoR1/(µL)	Xba1/(µL)	Spe1/(µL)	Pst1/(µL)	Buffer/(µL)		BSA/(µL)	MilliQ/(µL)	Total/(µL)
8/2 dddD PCR product	8.2	1	-	1	-	H	3	-	16.8	30
8/2 dddD PCR product control	0.41	-	-	-	-	H	1	-	8.6	10
pSB1C3	12.7	1	-	1	-	H	3	-	12.3	30
pSB1C3 control	0.04	-	-	-	-	H	1	-	8.4	10

8/3

@

Murata

Lane	Sample
1	pSB1C3-dddD(1)
2	pSB1C3-dddD(2)
3	pSB1C3-dddD(3)
4	1kb ladder (3µL)
5	1kb ladder (1µL)

Gel Extraction

Tatsui

Lane	Sample
1	1kb ladder
2	Blank
3	pSB1C3 2070bp
4	Blank
5	pSB1C3
6	Blank
7	8/2 dddD PCR product 2544bp
8	Blank
9	8/2 dddD PCR product

Sample	Concentration/(µg/mL)	260/280	260/230
pSB1C3	@	1.37	0.42
dddD PCR product	35	1.54	0.85

8/4

Colony PCR

Honda

Master Mix

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
40	1.6	1.6	36.8	80

Predenature	Denature	Annealing	Extension
94°C	94°C	55°C	68°C
2min	30sec	30sec	24sec

Denature-Annealing-Extension 30cycle

1	dddD+pSB1C3(1)-1
2	dddD+pSB1C3(1)-2
3	dddD+pSB1C3(2)-1
4	dddD+pSB1C3(2)-2
5	dddD+pSB1C3(2)-3
6	dddD+pSB1C3(2)-4
7	pSB1C3 control-1
8	control

Electrophoresis

Matsumoto

Lane	Sample
1	1kbp ladder
2	dddD+pSB1C3(1)-1
3	dddD+pSB1C3(1)-2
4	dddD+pSB1C3(2)-1
5	dddD+pSB1C3(2)-2
6	dddD+pSB1C3(2)-3
7	dddD+pSB1C3(2)-4
8	pSB1C3 control-1
9	control
10	1kbp ladder

PCR

Honda

Unexpected bands are confirmed in former Electrophoresis, so contamination of MilliQ or Quicktaq is suspected. For confirming contamination, PCR and Electrophoresis are conducted.

* suspicious samples for contamination

(1)

*Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
5	0.2	0.2	4.6	10

(2)

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
5	0.2	0.2	4.6	10

(3)

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	*MilliQ/(µL)	Total/(µL)
5	0.2	0.2	4.6	10

(4)

*Quick Tag/(µL)	VF2/(µL)	VR/(µL)	*MilliQ/(µL)	Total/(µL)
5	0.2	0.2	4.6	10

Predenature	Denature	Annealing	Extension
94°C	94°C	55°C	68°C
2min	30sec	30sec	24sec

Denature-Annealing-Extension 30cycle

Electrophoresis

Honda

Lane	Sample
1	1kbp ladder
2	(1)
3	(2)
4	(3)
5	(4)

8/18

PCR

Honda

For confirming contamination of VF2 and VR primers, we pour samples into new tubes and perform PCR and Electrophoresis.

(1)

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
5	0.2	0.2	4.6	10

control(2)

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	8/2 Pcon3/(µL)	MilliQ/(µL)	Total/(µL)
5	0.2	0.2	0.6	4.6	10

(3)

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
5	0.2	0.2	4.6	10

(4)

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	8/2 Pcon3/(µL)	MilliQ/(µL)	Total/(µL)
5	0.2	0.2	0.6	4.0	10

Predenature	Denature	Annealing	Extension
94°C	94°C	55°C	68°C
2min	30sec	30sec	24sec

Denature-Annealing-Extension 30cycle

Electrophoresis

Honda

Number	Sample
1	1kbp ladder
2	(1)
3	(2)
4	(3)
5	(4)

Restriction Enzyme Digestion

Honda

Sample/(µL)		EcoR1/(µL)	Xba1/(µL)	Spe1/(µL)	Pst1/(µL)	Buffer/(µL)		BSA/(µL)	MilliQ/(µL)	Total/(µL)
dddD 8/2 PCR product	5.5	0.7	-	0.7	-	H	2	-	11.1	20
dddD 8/2 PCR product control	0.41	-	-	-	-	H	1	-	8.6	10
pSB1C3	12.7	1	-	1	-	H	3	-	8.4	10
pSB1C3 control	0.64	-	-	-	-	H	1	-	8.4	10

Colony PCR

Honda

Master Mix

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
40	1.6	1.6	36.8	80

Predenature	Denature	Annealing	Extension
94°C	94°C	55°C	68°C
2min	30sec	30sec	24sec

Denature-Annealing-Extension 30cycle

Number	Sample
1	dddD+pSB1C3(1)-1
2	dddD+pSB1C3(1)-2
3	dddD+pSB1C3(2)-1
4	dddD+pSB1C3(2)-2
5	dddD+pSB1C3(2)-3
6	dddD+pSB1C3(2)-4
7	pSB1C3 control-1
8	control

Electrophoresis

Honda

Lane	Sample
1	1kbp ladder
2	dddD+pSB1C3(1)-1
3	dddD+pSB1C3(1)-2
4	dddD+pSB1C3(2)-1
5	dddD+pSB1C3(2)-2
6	dddD+pSB1C3(2)-3
7	dddD+pSB1C3(2)-4
8	pSB1C3 control-1
9	control

Gel Extraction

Murata

Lane	Sample
1	1kbp ladder
2	dddD control
3	dddD E,S
4	@
5	pSB1C3 control
6	pSB1C3 E,S

Sample	Concentration/(µg/mL)	260/280	260/230
105	1.30	0.85
dddD	105	1.27	0.73

8/19

Ligation

Honda,Li,Yamauchi

(1)

Vector/(µL)		Insert/(µL)		Buffer/(µL)		Ligase/(µL)		MilliQ/(µL)	Total/(µL)
pSB1C3	3.6	dddD	1	2xBuffer	5.6	T4Ligase	1	-	11.2

(2)

Vector/(µL)		Insert/(µL)		Buffer/(µL)		Ligase/(µL)		MilliQ/(µL)	Total/(µL)
pSB1C3	3.6	-	-	2xBuffer	5.6	T4Ligase	1	1	11.2

8/20

Transformation

Murata

Sample/(µL)		CompetentCells/(µL)		Total/(µL)	Medium
8/19 Ligation (1)	11.2	@	50	61.2	SOC
8/19 Ligation (2)	11.2	@	50	61.2	SOC

Ligation

Nakamura,Honda

(1)

Vector/(µL)		Insert/(µL)		Buffer/(µL)		Ligase/(µL)		MilliQ/(µL)	Total/(µL)
pSB1C3	10.8	dddD	3	2xBuffer	16.8	T4Ligase	3	-	33.6

(2)

Vector/(µL)		Insert/(µL)		Buffer/(µL)		Ligase/(µL)		MilliQ/(µL)	Total/(µL)
pSB1C3	10.8	-	-	2xBuffer	16.8	T4Ligase	3	3	33.6

Transformation

Nakamura,Honda

Sample/(µL)		CompetentCells/(µL)		Total/(µL)	Medium
dddD+pSB1C3	5	@	20	25	SOC
control	5	@	20	25	SOC

@

8/21

Colony PCR

Honda,Nakamura

Master Mix

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
35	1.4	1.4	32.2	70

Predenature	Denature	Annealing	Extension
94°C	94°C	55°C	68°C
2min	30sec	30sec	24sec

Denature-Annealing-Extension 30cycle

Electrophoresis

Nakamura

Number	Sample
1	8/20 dddD+pSB1C3(1)-1
2	8/20 dddD+pSB1C3(1)-2
3	8/20 dddD+pSB1C3(2)-1
4	8/20 dddD+pSB1C3(2)-2
5	8/20 dddD+pSB1C3(3)-1
6	8/20 dddD+pSB1C3(3)-2
7	control

Number	Sample
1	1kbp ladder
2	8/20 dddD+pSB1C3(1)-1
3	8/20 dddD+pSB1C3(1)-2
4	8/20 dddD+pSB1C3(2)-1
5	8/20 dddD+pSB1C3(2)-2
6	8/20 dddD+pSB1C3(3)-1
7	8/20 dddD+pSB1C3(3)-2
8	control

Gel Extraction

Yasuda

Lane	Sample
1	1kbp ladder
2	8/22 Mp(1) E,S
3	8/22 Mp(1) E,S
4	8/22 Mp(1) E,S

Lane	Sample
1	1kbp ladder
2	8/22 Mp(2) E,S
3	8/22 Mp(2) E,S
4	Blank
5	1kbp ladder
6	8/22 Mp(3) E,S
7	8/22 Mp(3) E,S

8/25

Gel Extraction

Yasuda

Lane	Sample
@	@

Ligation

Yasuda

Vector/(µL)		Insert/(µL)		Buffer/(µL)		Ligase/(µL)		MilliQ/(µL)	Total/(µL)
8/22 Mp(1) E,S	1.3	8/18 dddD gel extraction product	0.81	2x Buffer	5	T4 Ligase	0.3	2.69	10
8/22 Mp(2) E+S	0.95	8/18 dddD gel extraction product	0.71	2x Buffer	5	T4 Ligase	0.3	3.04	10
8/22 Mp(3) E+S	0.69	8/18 dddD gel extraction product	0.71	2x Buffer	5	T4Ligase	0.3	3.3	10

8/26

Transformation

Nakamura,Matsumoto

Sample/(µL)		CompetentCells/(µL)		Total/(µL)	Medium
8/25 Ligation (1)	10	@	50	60	SOC
8/25 Ligation(2)	10	@	50	60	SOC
8/25 Ligation(3)	10	@	30	40	SOC

@

8/27

Colony PCR

Murata

Master Mix

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
80	3.2	3.2	73.6	160

Predenature	Denature	Annealing	Extension
94°C	94°C	55°C	68°C
2min	30sec	30sec	2h20min

Denature-Annealing-Extension 30cycle

Number	Sample
1	8/26 dddD+pSB1C3(1)-1
2	8/26 dddD+pSB1C3(1)-2
3	8/26 dddD+pSB1C3(1)-3
4	8/26 dddD+pSB1C3(1)-4
5	8/26 dddD+pSB1C3(1)-5
6	8/26 dddD+pSB1C3(2)-1
7	8/26 dddD+pSB1C3(2)-2
8	8/26 dddD+pSB1C3(2)-3
9	8/26 dddD+pSB1C3(2)-4
10	8/26 dddD+pSB1C3(2)-5
11	8/26 dddD+pSB1C3(3)-1
12	8/26 dddD+pSB1C3(3)-2
13	8/26 dddD+pSB1C3(3)-3
14	8/26 dddD+pSB1C3(3)-4
15	8/26 dddD+pSB1C3(3)-5
16	control

Electrophoresis

Yasuda

Lane	Sample
1	1kbp ladder
2	8/26 dddD+pSB1C3(1)-1
3	8/26 dddD+pSB1C3(1)-2
4	8/26 dddD+pSB1C3(1)-3
5	8/26 dddD+pSB1C3(1)-4
6	8/26 dddD+pSB1C3(1)-5

Lane	Sample
1	1kbp ladder
2	8/26 dddD+pSB1C3(2)-1
3	8/26 dddD+pSB1C3(2)-2
4	8/26 dddD+pSB1C3(2)-3
5	8/26 dddD+pSB1C3(2)-4
6	8/26 dddD+pSB1C3(2)-5
7	1kbp ladder
8	8/26 dddD+pSB1C3(3)-1
9	8/26 dddD+pSB1C3(3)-2
10	8/26 dddD+pSB1C3(3)-3
11	8/26 dddD+pSB1C3(3)-4
12	8/26 dddD+pSB1C3(3)-5
13	1kbp ladder
14	control

8/29

PCR

Matsumoto

Master Mix

Quick Tag/(µL)	VF2/(µL)	VR/(µL)	MilliQ/(µL)	Total/(µL)
@	@	@	@	25

Predenature	Denature	Annealing	Extension
95°C	98°C	65°C	72°C
5min	20sec	15sec	50sec

Denature-Annealing-Extension @cycle

DNA Purification

Matsumoto

Sample	Concentration/(µg/mL)	260/280	260/230
8/29 Pre-dddD-Suf PCR product	335	1.73	1.91

Restriction Enzyme Digestion

Matsumoto

Sample/(µL)		EcoR1/(µL)	Xba1/(µL)	Spe1/(µL)	Pst1/(µL)	Buffer/(µL)		BSA/(µL)	MilliQ/(µL)	Total/(µL)
8/29 Pre-dddD-Suf PCR product	6.0	1	-	1	-	H	3	-	19	30

DNA Purification

Matsumoto

Sample	Concentration/(µg/mL)	260/280	260/230
8/29 Pre-dddD-Suf Restriction Enzyme Digestion product	60	1.64	0.94

Ligation

Matsumoto

Vector/(µL)		Insert/(µL)		Buffer/(µL)		Ligase/(µL)		MilliQ/(µL)	Total/(µL)
pSB1C3 E,S gel extraction product	4.7	dddD-pSB1C3	4	2x Buffer	9.7	T4 Ligase	1	-	19.4
pSB1C3 E,S gel extraction product	4.7	-	-	2x Buffer	9.7	T4 Ligase	1	4	19.4

Transformation

Matsumoto

Sample/(µL)		CompetentCells/(µL)		Total/(µL)	Medium
dddD 8/29 ligation product	10	DH5a	40	50	SOC
8/29 ligation product control	10	DH5a	40	50	SOC
pSB1C3 control	1	DH5a	20	@	SOC

@@ Line 2: / Line 2: @@
     <div id="kyoto-notebook">
 <!-- 警告：ここより上を消したら怒る -->
+<div>
+	<a name="0731" class="kyoto-jump"></a>
+	<h3>7/31</h3>
+		<h4>Restriction Enzyme Digestion</h4>
+			<span class="kyoto-author">Nakamura and Matsumoto</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th>EcoR1/(&micro;L)</th>
+					<th>Xba1/(&micro;L)</th>
+					<th>Spe1/(&micro;L)</th>
+					<th>Pst1/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th>BSA/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>3/24 dddD PCR product</td>
+					<td>3.2</td>
+					<td>1</td>
+					<td>-</td>
+					<td>1</td>
+					<td>-</td>
+					<td>H</td>
+					<td>3</td>
+					<td>-</td>
+					<td>21.8</td>
+					<td>30</td>
+				</tr>
+				<tr>
+					<td>pSB1C3</td>
+					<td>18.5</td>
+					<td>1</td>
+					<td>-</td>
+					<td>1</td>
+					<td>-</td>
+					<td>H</td>
+					<td>3</td>
+					<td>-</td>
+					<td>6.5</td>
+					<td>30</td>
+				</tr>
+				<tr>
+					<td>pSB1C3 control</td>
+					<td>6.2</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>H</td>
+					<td>1</td>
+					<td>-</td>
+					<td>2.8</td>
+					<td>10</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0801" class="kyoto-jump"></a>
+	<h3>8/1</h3>
+		<h4>Gel Extraction</h4>
+			<span class="kyoto-author">Murata and Yasuda</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>3/24 dddD PCR product E,S</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>3/24 dddD PCR product E,S</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>Blank</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>pSB1C3 E,S</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>pSB1C3 E,S</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>Blank</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>9</td>
+					<td>3/24 dddD PCR product control</td>
+				<tr>
+					<td>10</td>
+					<td>pSB1C3 control</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Sample</th>
+					<th>Concentration/(&micro;g/mL)</th>
+					<th>260/280</th>
+					<th>260/230</th>
+				</tr>
+				<tr>
+					<td>PSB1C3</td>
+					<td>13.3</td>
+					<td>1.90</td>
+					<td>-</td>
+				</tr>
+				<tr>
+					<td>dddD</td>
+					<td>10.8</td>
+					<td>1.64</td>
+					<td>0.15</td>
+				</tr>
+			</table>
+		<h4>Ligation</h4>
+			<span class="kyoto-author">Yasuda</span>
+			<table>
+				<tr>
+					<th colspan=2>Vector/(&micro;L)</th>
+					<th colspan=2>Insert/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th colspan=2>Ligase/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</tr>
+				<tr>
+					<td>8/1 PSB1C3 gel extraction product 2047bp</td>
+					<td>1.5</td>
+					<td>8/1 dddD gel extraction product 2545bp</td>
+					<td>6.9</td>
+					<td>10xT4Ligase</td>
+					<td>2</td>
+					<td>T4Ligase</td>
+					<td>1</td>
+					<td>8.6</td>
+					<td>20</td>
+				</tr>
+			</table>
+		<h4>Transformation</h4>
+			<span class="kyoto-author">Murata</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th colspan=2>CompetentCells/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+					<th>Medium</th>
+				</tr>
+				<tr>
+					<td>PSB1C3-dddD</td>
+					<td>3</td>
+					<td>コンピ名</td>
+					<td>30</td>
+					<td>33</td>
+					<td>SOC</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0802" class="kyoto-jump"></a>
+	<h3>8/2</h3>
+		<h4>PCR Cloning</h4>
+			<span class="kyoto-author">Murata</span>
+			<table>
+				<tr>
+					<th>dddD without Pre-Suf</th>
+					<th>2x KAPA HiFi Hotstart Ready Mix/(&micro;L)</th>
+					<th>F Pre-dddD PCR product 62/(&micro;L)</th>
+					<th>Rv Spe1-dddD PCR product 68/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>pick up</td>
+					<td>12.5</td>
+					<td>0.75</td>
+					<td>0.75</td>
+					<td>11</td>
+					<td>25</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>95&deg;C</td>
+					<td>98&deg;C</td>
+					<td>65&deg;C</td>
+					<td>72&deg;C</td>
+				</tr>
+				<tr>
+					<td>5min</td>
+					<td>20sec</td>
+					<td>15sec</td>
+					<td>50sec</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension 25cycle</p>
+		<h4>DNA Purification</h4>
+			<span class="kyoto-author">Murata</span>
+			<p>8/2 Pre-dddD-Suf PCR product</p>
+		<h4>Colony PCR</h4>
+			<span class="kyoto-author">Honda,Nakamura and Matsumoto</span>
+			<p>Master Mix</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>15</td>
+					<td>0.6</td>
+					<td>0.6</td>
+					<td>13.8</td>
+					<td>30</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>94&deg;C</td>
+					<td>94&deg;C</td>
+					<td>55&deg;C</td>
+					<td>68&deg;C</td>
+				</tr>
+				<tr>
+					<td>2min</td>
+					<td>30sec</td>
+					<td>30sec</td>
+					<td>24sec</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension 30cycle</p>
+			<table>
+				<tr>
+					<th>Number</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>pSB1C3-dddD(1)</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>pSB1C3-dddD(2)</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>pSB1C3-dddD(3)</td>
+				</tr>
+			</table>
+		<h4>Concentration Measurement</h4>
+			<span class="kyoto-author">Nakamura and Matsumoto</span>
+			<table>
+				<tr>
+					<th>Sample</th>
+					<th>Concentration/(&micro;g/mL)</th>
+					<th>260/280</th>
+					<th>260/230</th>
+				</tr>
+				<tr>
+					<td>dddD PCR product</td>
+					<td>245</td>
+					<td>1.59</td>
+					<td>2.17</td>
+				</tr>
+			</table>
+		<h4>Restriction Enzyme Digestion</h4>
+			<span class="kyoto-author">Nakamura and Matsumoto</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th>EcoR1/(&micro;L)</th>
+					<th>Xba1/(&micro;L)</th>
+					<th>Spe1/(&micro;L)</th>
+					<th>Pst1/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th>BSA/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>8/2 dddD PCR product</td>
+					<td>8.2</td>
+					<td>1</td>
+					<td>-</td>
+					<td>1</td>
+					<td>-</td>
+					<td>H</td>
+					<td>3</td>
+					<td>-</td>
+					<td>16.8</td>
+					<td>30</td>
+				</tr>
+				<tr>
+					<td>8/2 dddD PCR product control</td>
+					<td>0.41</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>H</td>
+					<td>1</td>
+					<td>-</td>
+					<td>8.6</td>
+					<td>10</td>
+				</tr>
+				<tr>
+					<td>pSB1C3</td>
+					<td>12.7</td>
+					<td>1</td>
+					<td>-</td>
+					<td>1</td>
+					<td>-</td>
+					<td>H</td>
+					<td>3</td>
+					<td>-</td>
+					<td>12.3</td>
+					<td>30</td>
+				</tr>
+				<tr>
+					<td>pSB1C3 control</td>
+					<td>0.04</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>H</td>
+					<td>1</td>
+					<td>-</td>
+					<td>8.4</td>
+					<td>10</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0803" class="kyoto-jump"></a>
+	<h3>8/3</h3>
+		<h4>@</h4>
+			<span class="kyoto-author">Murata</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>pSB1C3-dddD(1)</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>pSB1C3-dddD(2)</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>pSB1C3-dddD(3)</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>1kb ladder (3&micro;L)</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>1kb ladder (1&micro;L)</td>
+				</tr>
+			</table>
+		<h4>Gel Extraction</h4>
+			<span class="kyoto-author">Tatsui</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kb ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>Blank</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>pSB1C3 2070bp</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>Blank</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>pSB1C3</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>Blank</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>8/2 dddD PCR product 2544bp</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>Blank</td>
+				</tr>
+				<tr>
+					<td>9</td>
+					<td>8/2 dddD PCR product</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Sample</th>
+					<th>Concentration/(&micro;g/mL)</th>
+					<th>260/280</th>
+					<th>260/230</th>
+				</tr>
+				<tr>
+					<td>pSB1C3</td>
+					<td>@</td>
+					<td>1.37</td>
+					<td>0.42</td>
+				</tr>
+				<tr>
+					<td>dddD PCR product</td>
+					<td>35</td>
+					<td>1.54</td>
+					<td>0.85</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0804" class="kyoto-jump"></a>
+	<h3>8/4</h3>
+		<h4>Colony PCR</h4>
+			<span class="kyoto-author">Honda</span>
+			<p>Master Mix</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>40</td>
+					<td>1.6</td>
+					<td>1.6</td>
+					<td>36.8</td>
+					<td>80</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>94&deg;C</td>
+					<td>94&deg;C</td>
+					<td>55&deg;C</td>
+					<td>68&deg;C</td>
+				</tr>
+				<tr>
+					<td>2min</td>
+					<td>30sec</td>
+					<td>30sec</td>
+					<td>24sec</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension 30cycle</p>
+			<table>
+				<tr>
+					<th>1</th>
+					<th>dddD+pSB1C3(1)-1</th>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>dddD+pSB1C3(1)-2</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>dddD+pSB1C3(2)-1</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>dddD+pSB1C3(2)-2</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>dddD+pSB1C3(2)-3</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>dddD+pSB1C3(2)-4</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>pSB1C3 control-1</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>control</td>
+				</tr>
+			</table>
+		<h4>Electrophoresis</h4>
+			<span class="kyoto-author">Matsumoto</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>dddD+pSB1C3(1)-1</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>dddD+pSB1C3(1)-2</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>dddD+pSB1C3(2)-1</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>dddD+pSB1C3(2)-2</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>dddD+pSB1C3(2)-3</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>dddD+pSB1C3(2)-4</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>pSB1C3 control-1</td>
+				</tr>
+				<tr>
+					<td>9</td>
+					<td>control</td>
+				</tr>
+				<tr>
+					<td>10</td>
+					<td>1kbp ladder</td>
+				</tr>
+			</table>
+		<h4>PCR</h4>
+			<span class="kyoto-author">Honda</span>
+			<p>Unexpected bands are  confirmed in former Electrophoresis, so contamination of MilliQ or Quicktaq is suspected. For confirming contamination, PCR and Electrophoresis are conducted. </p>
+			<p>* suspicious samples for contamination</p>
+			<p>(1)</p>
+			<table>
+				<tr>
+					<th>*Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>0.2</td>
+					<td>0.2</td>
+					<td>4.6</td>
+					<td>10</td>
+				</tr>
+			</table>
+			<p>(2)</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>0.2</td>
+					<td>0.2</td>
+					<td>4.6</td>
+					<td>10</td>
+				</tr>
+			</table>
+			<p>(3)</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>*MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>0.2</td>
+					<td>0.2</td>
+					<td>4.6</td>
+					<td>10</td>
+				</tr>
+			</table>
+			<p>(4)</p>
+			<table>
+				<tr>
+					<th>*Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>*MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>0.2</td>
+					<td>0.2</td>
+					<td>4.6</td>
+					<td>10</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>94&deg;C</td>
+					<td>94&deg;C</td>
+					<td>55&deg;C</td>
+					<td>68&deg;C</td>
+				</tr>
+				<tr>
+					<td>2min</td>
+					<td>30sec</td>
+					<td>30sec</td>
+					<td>24sec</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension 30cycle</p>
+		<h4>Electrophoresis</h4>
+			<span class="kyoto-author">Honda</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>(1)</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>(2)</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>(3)</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>(4)</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0818" class="kyoto-jump"></a>
+	<h3>8/18</h3>
+		<h4>PCR</h4>
+			<span class="kyoto-author">Honda</span>
+			<p>For confirming contamination of VF2 and VR primers, we pour samples into new tubes and perform PCR and Electrophoresis.</p>
+			<p>(1)</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>0.2</td>
+					<td>0.2</td>
+					<td>4.6</td>
+					<td>10</td>
+				</tr>
+			</table>
+			<p>control(2)</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>8/2 Pcon3/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>0.2</td>
+					<td>0.2</td>
+					<td>0.6</td>
+					<td>4.6</td>
+					<td>10</td>
+				</tr>
+			</table>
+			<p>(3)</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>0.2</td>
+					<td>0.2</td>
+					<td>4.6</td>
+					<td>10</td>
+				</tr>
+			</table>
+			<p>(4)</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>8/2 Pcon3/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>0.2</td>
+					<td>0.2</td>
+					<td>0.6</td>
+					<td>4.0</td>
+					<td>10</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>94&deg;C</td>
+					<td>94&deg;C</td>
+					<td>55&deg;C</td>
+					<td>68&deg;C</td>
+				</tr>
+				<tr>
+					<td>2min</td>
+					<td>30sec</td>
+					<td>30sec</td>
+					<td>24sec</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension 30cycle</p>
+		<h4>Electrophoresis</h4>
+			<span class="kyoto-author">Honda</span>
+			<table>
+				<tr>
+					<th>Number</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>(1)</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>(2)</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>(3)</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>(4)</td>
+				</tr>
+			</table>
+		<h4>Restriction Enzyme Digestion</h4>
+			<span class="kyoto-author">Honda</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th>EcoR1/(&micro;L)</th>
+					<th>Xba1/(&micro;L)</th>
+					<th>Spe1/(&micro;L)</th>
+					<th>Pst1/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th>BSA/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>dddD 8/2 PCR product</td>
+					<td>5.5</td>
+					<td>0.7</td>
+					<td>-</td>
+					<td>0.7</td>
+					<td>-</td>
+					<td>H</td>
+					<td>2</td>
+					<td>-</td>
+					<td>11.1</td>
+					<td>20</td>
+				</tr>
+				<tr>
+					<td>dddD 8/2 PCR product control</td>
+					<td>0.41</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>H</td>
+					<td>1</td>
+					<td>-</td>
+					<td>8.6</td>
+					<td>10</td>
+				</tr>
+				<tr>
+					<td>pSB1C3</td>
+					<td>12.7</td>
+					<td>1</td>
+					<td>-</td>
+					<td>1</td>
+					<td>-</td>
+					<td>H</td>
+					<td>3</td>
+					<td>-</td>
+					<td>8.4</td>
+					<td>10</td>
+				</tr>
+				<tr>
+					<td>pSB1C3 control</td>
+					<td>0.64</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>-</td>
+					<td>H</td>
+					<td>1</td>
+					<td>-</td>
+					<td>8.4</td>
+					<td>10</td>
+				</tr>
+			</table>
+		<h4>Colony PCR</h4>
+			<span class="kyoto-author">Honda</span>
+			<p>Master Mix</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>40</td>
+					<td>1.6</td>
+					<td>1.6</td>
+					<td>36.8</td>
+					<td>80</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>94&deg;C</td>
+					<td>94&deg;C</td>
+					<td>55&deg;C</td>
+					<td>68&deg;C</td>
+				</tr>
+				<tr>
+					<td>2min</td>
+					<td>30sec</td>
+					<td>30sec</td>
+					<td>24sec</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension 30cycle</p>
+			<table>
+				<tr>
+					<th>Number</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>dddD+pSB1C3(1)-1</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>dddD+pSB1C3(1)-2</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>dddD+pSB1C3(2)-1</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>dddD+pSB1C3(2)-2</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>dddD+pSB1C3(2)-3</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>dddD+pSB1C3(2)-4</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>pSB1C3 control-1</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>control</td>
+				</tr>
+			</table>
+		<h4>Electrophoresis</h4>
+			<span class="kyoto-author">Honda</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>dddD+pSB1C3(1)-1</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>dddD+pSB1C3(1)-2</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>dddD+pSB1C3(2)-1</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>dddD+pSB1C3(2)-2</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>dddD+pSB1C3(2)-3</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>dddD+pSB1C3(2)-4</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>pSB1C3 control-1</td>
+				</tr>
+				<tr>
+					<td>9</td>
+					<td>control</td>
+				</tr>
+			</table>
+		<h4>Gel Extraction</h4>
+			<span class="kyoto-author">Murata</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>dddD control</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>dddD E,S</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>@</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>pSB1C3 control</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>pSB1C3 E,S</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Sample</th>
+					<th>Concentration/(&micro;g/mL)</th>
+					<th>260/280</th>
+					<th>260/230</th>
+				</tr>
+				<tr>
+					<tdpSB1C3</td>
+					<td>105</td>
+					<td>1.30</td>
+					<td>0.85</td>
+				</tr>
+				<tr>
+					<td>dddD</td>
+					<td>105</td>
+					<td>1.27</td>
+					<td>0.73</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0819" class="kyoto-jump"></a>
+	<h3>8/19</h3>
+		<h4>Ligation</h4>
+			<span class="kyoto-author">Honda,Li,Yamauchi</span>
+			<p>(1)</p>
+			<table>
+				<tr>
+					<th colspan=2>Vector/(&micro;L)</th>
+					<th colspan=2>Insert/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th colspan=2>Ligase/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</tr>
+				<tr>
+					<td>pSB1C3</td>
+					<td>3.6</td>
+					<td>dddD</td>
+					<td>1</td>
+					<td>2xBuffer</td>
+					<td>5.6</td>
+					<td>T4Ligase</td>
+					<td>1</td>
+					<td>-</td>
+					<td>11.2</td>
+				</tr>
+			</table>
+			<p>(2)</p>
+			<table>
+				<tr>
+					<th colspan=2>Vector/(&micro;L)</th>
+					<th colspan=2>Insert/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th colspan=2>Ligase/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</tr>
+				<tr>
+					<td>pSB1C3</td>
+					<td>3.6</td>
+					<td>-</td>
+					<td>-</td>
+					<td>2xBuffer</td>
+					<td>5.6</td>
+					<td>T4Ligase</td>
+					<td>1</td>
+					<td>1</td>
+					<td>11.2</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0820" class="kyoto-jump"></a>
+	<h3>8/20</h3>
+		<h4>Transformation</h4>
+			<span class="kyoto-author">Murata</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th colspan=2>CompetentCells/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+					<th>Medium</th>
+				</tr>
+				<tr>
+					<td>8/19 Ligation (1)</td>
+					<td>11.2</td>
+					<td>@</td>
+					<td>50</td>
+					<td>61.2</td>
+					<td>SOC</td>
+				</tr>
+				<tr>
+					<td>8/19 Ligation (2)</td>
+					<td>11.2</td>
+					<td>@</td>
+					<td>50</td>
+					<td>61.2</td>
+					<td>SOC</td>
+				</tr>
+			</table>
+		<h4>Ligation</h4>
+			<span class="kyoto-author">Nakamura,Honda</span>
+			<p>(1)</p>
+			<table>
+				<tr>
+					<th colspan=2>Vector/(&micro;L)</th>
+					<th colspan=2>Insert/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th colspan=2>Ligase/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</tr>
+				<tr>
+					<td>pSB1C3</td>
+					<td>10.8</td>
+					<td>dddD</td>
+					<td>3</td>
+					<td>2xBuffer</td>
+					<td>16.8</td>
+					<td>T4Ligase</td>
+					<td>3</td>
+					<td>-</td>
+					<td>33.6</td>
+				</tr>
+			</table>
+			<p>(2)</p>
+			<table>
+				<tr>
+					<th colspan=2>Vector/(&micro;L)</th>
+					<th colspan=2>Insert/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th colspan=2>Ligase/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</tr>
+				<tr>
+					<td>pSB1C3</td>
+					<td>10.8</td>
+					<td>-</td>
+					<td>-</td>
+					<td>2xBuffer</td>
+					<td>16.8</td>
+					<td>T4Ligase</td>
+					<td>3</td>
+					<td>3</td>
+					<td>33.6</td>
+				</tr>
+			</table>
+		<h4>Transformation</h4>
+			<span class="kyoto-author">Nakamura,Honda</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th colspan=2>CompetentCells/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+					<th>Medium</th>
+				</tr>
+				<tr>
+					<td>dddD+pSB1C3</td>
+					<td>5</td>
+					<td>@</td>
+					<td>20</td>
+					<td>25</td>
+					<td>SOC</td>
+				</tr>
+				<tr>
+					<td>control</td>
+					<td>5</td>
+					<td>@</td>
+					<td>20</td>
+					<td>25</td>
+					<td>SOC</td>
+				</tr>
+			</table>
+			<p>@</P>
+</div>
+<div>
+	<a name="0821" class="kyoto-jump"></a>
+	<h3>8/21</h3>
+		<h4>Colony PCR</h4>
+			<span class="kyoto-author">Honda,Nakamura</span>
+			<p>Master Mix</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>35</td>
+					<td>1.4</td>
+					<td>1.4</td>
+					<td>32.2</td>
+					<td>70</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>94&deg;C</td>
+					<td>94&deg;C</td>
+					<td>55&deg;C</td>
+					<td>68&deg;C</td>
+				</tr>
+				<tr>
+					<td>2min</td>
+					<td>30sec</td>
+					<td>30sec</td>
+					<td>24sec</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension 30cycle</p>
+			<table>
+				<tr>
+					<th>Number</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>8/20 dddD+pSB1C3(1)-1</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>8/20 dddD+pSB1C3(1)-2</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>8/20 dddD+pSB1C3(2)-1</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>8/20 dddD+pSB1C3(2)-2</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>8/20 dddD+pSB1C3(3)-1</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>8/20 dddD+pSB1C3(3)-2</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>control</td>
+				</tr>
+			<h4>Electrophoresis</h4>
+			<span class="kyoto-author">Nakamura</span>
+				<table>
+				<tr>
+					<th>Number</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>8/20 dddD+pSB1C3(1)-1</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>8/20 dddD+pSB1C3(1)-2</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>8/20 dddD+pSB1C3(2)-1</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>8/20 dddD+pSB1C3(2)-2</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>8/20 dddD+pSB1C3(3)-1</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>8/20 dddD+pSB1C3(3)-2</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>control</td>
+				</tr>
+			</table>
+			<h4>Gel Extraction</h4>
+			<span class="kyoto-author">Yasuda</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>8/22 Mp(1) E,S</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>8/22 Mp(1) E,S</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>8/22 Mp(1) E,S</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>8/22 Mp(2) E,S</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>8/22 Mp(2) E,S</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>Blank</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>8/22 Mp(3) E,S</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>8/22 Mp(3) E,S</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0825" class="kyoto-jump"></a>
+	<h3>8/25</h3>
+		<h4>Gel Extraction</h4>
+			<span class="kyoto-author">Yasuda</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>@</td>
+					<td>@</td>
+				</tr>
+			</table>
+		<h4>Ligation</h4>
+			<span class="kyoto-author">Yasuda</span>
+			<table>
+				<tr>
+					<th colspan=2>Vector/(&micro;L)</th>
+					<th colspan=2>Insert/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th colspan=2>Ligase/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</tr>
+				<tr>
+					<td>8/22 Mp(1) E,S</td>
+					<td>1.3</td>
+					<td>8/18 dddD gel extraction product</td>
+					<td>0.81</td>
+					<td>2x Buffer</td>
+					<td>5</td>
+					<td>T4 Ligase</td>
+					<td>0.3</td>
+					<td>2.69</td>
+					<td>10</td>
+				</tr>
+				<tr>
+					<td>8/22 Mp(2) E+S</td>
+					<td>0.95</td>
+					<td>8/18 dddD gel extraction product</td>
+					<td>0.71</td>
+					<td>2x Buffer</td>
+					<td>5</td>
+					<td>T4 Ligase</td>
+					<td>0.3</td>
+					<td>3.04</td>
+					<td>10</td>
+				</tr>
+				<tr>
+					<td>8/22 Mp(3) E+S</td>
+					<td>0.69</td>
+					<td>8/18 dddD gel extraction product</td>
+					<td>0.71</td>
+					<td>2x Buffer</td>
+					<td>5</td>
+					<td>T4Ligase</td>
+					<td>0.3</td>
+					<td>3.3</td>
+					<td>10</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0826" class="kyoto-jump"></a>
+	<h3>8/26</h3>
+		<h4>Transformation</h4>
+			<span class="kyoto-author">Nakamura,Matsumoto</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th colspan=2>CompetentCells/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+					<th>Medium</th>
+				</tr>
+				<tr>
+					<td>8/25 Ligation (1)</td>
+					<td>10</td>
+					<td>@</td>
+					<td>50</td>
+					<td>60</td>
+					<td>SOC</td>
+				</tr>
+				<tr>
+					<td>8/25 Ligation(2)</td>
+					<td>10</td>
+					<td>@</td>
+					<td>50</td>
+					<td>60</td>
+					<td>SOC</td>
+				</tr>
+				<tr>
+					<td>8/25 Ligation(3)</td>
+					<td>10</td>
+					<td>@</td>
+					<td>30</td>
+					<td>40</td>
+					<td>SOC</td>
+				</tr>
+			</table>
+			<p>@</p>
+</div>
+<div>
+	<a name="0827" class="kyoto-jump"></a>
+	<h3>8/27</h3>
+		<h4>Colony PCR</h4>
+			<span class="kyoto-author">Murata</span>
+			<p>Master Mix</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>80</td>
+					<td>3.2</td>
+					<td>3.2</td>
+					<td>73.6</td>
+					<td>160</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>94&deg;C</td>
+					<td>94&deg;C</td>
+					<td>55&deg;C</td>
+					<td>68&deg;C</td>
+				</tr>
+				<tr>
+					<td>2min</td>
+					<td>30sec</td>
+					<td>30sec</td>
+					<td>2h20min</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension 30cycle</p>
+			<table>
+				<tr>
+					<th>Number</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>8/26 dddD+pSB1C3(1)-1</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>8/26 dddD+pSB1C3(1)-2</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>8/26 dddD+pSB1C3(1)-3</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>8/26 dddD+pSB1C3(1)-4</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>8/26 dddD+pSB1C3(1)-5</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>8/26 dddD+pSB1C3(2)-1</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>8/26 dddD+pSB1C3(2)-2</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>8/26 dddD+pSB1C3(2)-3</td>
+				</tr>
+				<tr>
+					<td>9</td>
+					<td>8/26 dddD+pSB1C3(2)-4</td>
+				</tr>
+				<tr>
+					<td>10</td>
+					<td>8/26 dddD+pSB1C3(2)-5</td>
+				</tr>
+				<tr>
+					<td>11</td>
+					<td>8/26 dddD+pSB1C3(3)-1</td>
+				</tr>
+				<tr>
+					<td>12</td>
+					<td>8/26 dddD+pSB1C3(3)-2</td>
+				</tr>
+				<tr>
+					<td>13</td>
+					<td>8/26 dddD+pSB1C3(3)-3</td>
+				</tr>
+				<tr>
+					<td>14</td>
+					<td>8/26 dddD+pSB1C3(3)-4</td>
+				</tr>
+				<tr>
+					<td>15</td>
+					<td>8/26 dddD+pSB1C3(3)-5</td>
+				</tr>
+				<tr>
+					<td>16</td>
+					<td>control</td>
+				</tr>
+			</table>
+		<h4>Electrophoresis</h4>
+			<span class="kyoto-author">Yasuda</span>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>8/26 dddD+pSB1C3(1)-1</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>8/26 dddD+pSB1C3(1)-2</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>8/26 dddD+pSB1C3(1)-3</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>8/26 dddD+pSB1C3(1)-4</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>8/26 dddD+pSB1C3(1)-5</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Lane</th>
+					<th>Sample</th>
+				</tr>
+				<tr>
+					<td>1</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>2</td>
+					<td>8/26 dddD+pSB1C3(2)-1</td>
+				</tr>
+				<tr>
+					<td>3</td>
+					<td>8/26 dddD+pSB1C3(2)-2</td>
+				</tr>
+				<tr>
+					<td>4</td>
+					<td>8/26 dddD+pSB1C3(2)-3</td>
+				</tr>
+				<tr>
+					<td>5</td>
+					<td>8/26 dddD+pSB1C3(2)-4</td>
+				</tr>
+				<tr>
+					<td>6</td>
+					<td>8/26 dddD+pSB1C3(2)-5</td>
+				</tr>
+				<tr>
+					<td>7</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>8</td>
+					<td>8/26 dddD+pSB1C3(3)-1</td>
+				</tr>
+				<tr>
+					<td>9</td>
+					<td>8/26 dddD+pSB1C3(3)-2</td>
+				</tr>
+				<tr>
+					<td>10</td>
+					<td>8/26 dddD+pSB1C3(3)-3</td>
+				</tr>
+				<tr>
+					<td>11</td>
+					<td>8/26 dddD+pSB1C3(3)-4</td>
+				</tr>
+				<tr>
+					<td>12</td>
+					<td>8/26 dddD+pSB1C3(3)-5</td>
+				</tr>
+				<tr>
+					<td>13</td>
+					<td>1kbp ladder</td>
+				</tr>
+				<tr>
+					<td>14</td>
+					<td>control</td>
+				</tr>
+			</table>
+</div>
+<div>
+	<a name="0829" class="kyoto-jump"></a>
+	<h3>8/29</h3>
+		<h4>PCR</h4>
+			<span class="kyoto-author">Matsumoto</span>
+			<p>Master Mix</p>
+			<table>
+				<tr>
+					<th>Quick Tag/(&micro;L)</th>
+					<th>VF2/(&micro;L)</th>
+					<th>VR/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>@</td>
+					<td>@</td>
+					<td>@</td>
+					<td>@</td>
+					<td>25</td>
+				</tr>
+			</table>
+			<table>
+				<tr>
+					<th>Predenature</th>
+					<th>Denature</th>
+					<th>Annealing</th>
+					<th>Extension</th>
+				</tr>
+				<tr>
+					<td>95&deg;C</td>
+					<td>98&deg;C</td>
+					<td>65&deg;C</td>
+					<td>72&deg;C</td>
+				</tr>
+				<tr>
+					<td>5min</td>
+					<td>20sec</td>
+					<td>15sec</td>
+					<td>50sec</td>
+				</tr>
+			</table>
+			<p>Denature-Annealing-Extension @cycle</p>
+		<h4>DNA Purification</h4>
+			<span class="kyoto-author">Matsumoto</span>
+			<table>
+				<tr>
+					<th>Sample</th>
+					<th>Concentration/(&micro;g/mL)</th>
+					<th>260/280</th>
+					<th>260/230</th>
+				</tr>
+				<tr>
+					<td>8/29 Pre-dddD-Suf PCR product</td>
+					<td>335</td>
+					<td>1.73</td>
+					<td>1.91</td>
+				</tr>
+			</table>
+		<h4>Restriction Enzyme Digestion</h4>
+			<span class="kyoto-author">Matsumoto</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th>EcoR1/(&micro;L)</th>
+					<th>Xba1/(&micro;L)</th>
+					<th>Spe1/(&micro;L)</th>
+					<th>Pst1/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th>BSA/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+				</tr>
+				<tr>
+					<td>8/29 Pre-dddD-Suf PCR product</td>
+					<td>6.0</td>
+					<td>1</td>
+					<td>-</td>
+					<td>1</td>
+					<td>-</td>
+					<td>H</td>
+					<td>3</td>
+					<td>-</td>
+					<td>19</td>
+					<td>30</td>
+				</tr>
+			</table>
+		<h4>DNA Purification</h4>
+			<span class="kyoto-author">Matsumoto</span>
+			<table>
+				<tr>
+					<th>Sample</th>
+					<th>Concentration/(&micro;g/mL)</th>
+					<th>260/280</th>
+					<th>260/230</th>
+				</tr>
+				<tr>
+					<td>8/29 Pre-dddD-Suf Restriction Enzyme Digestion product</td>
+					<td>60</td>
+					<td>1.64</td>
+					<td>0.94</td>
+				</tr>
+			</table>
+		<h4>Ligation</h4>
+			<span class="kyoto-author">Matsumoto</span>
+			<table>
+				<tr>
+					<th colspan=2>Vector/(&micro;L)</th>
+					<th colspan=2>Insert/(&micro;L)</th>
+					<th colspan=2>Buffer/(&micro;L)</th>
+					<th colspan=2>Ligase/(&micro;L)</th>
+					<th>MilliQ/(&micro;L)</th>
+					<th>Total/(&micro;L)</tr>
+				<tr>
+					<td>pSB1C3 E,S gel extraction product</td>
+					<td>4.7</td>
+					<td>dddD-pSB1C3</td>
+					<td>4</td>
+					<td>2x Buffer</td>
+					<td>9.7</td>
+					<td>T4 Ligase</td>
+					<td>1</td>
+					<td>-</td>
+					<td>19.4</td>
+				</tr>
+				<tr>
+					<td>pSB1C3 E,S gel extraction product</td>
+					<td>4.7</td>
+					<td>-</td>
+					<td>-</td>
+					<td>2x Buffer</td>
+					<td>9.7</td>
+					<td>T4 Ligase</td>
+					<td>1</td>
+					<td>4</td>
+					<td>19.4</td>
+				</tr>
+			</table>
+		<h4>Transformation</h4>
+			<span class="kyoto-author">Matsumoto</span>
+			<table>
+				<tr>
+					<th colspan=2>Sample/(&micro;L)</th>
+					<th colspan=2>CompetentCells/(&micro;L)</th>
+					<th>Total/(&micro;L)</th>
+					<th>Medium</th>
+				</tr>
+				<tr>
+					<td>dddD 8/29 ligation product</td>
+					<td>10</td>
+					<td>DH5a</td>
+					<td>40</td>
+					<td>50</td>
+					<td>SOC</td>
+				</tr>
+				<tr>
+					<td>8/29 ligation product control</td>
+					<td>10</td>
+					<td>DH5a</td>
+					<td>40</td>
+					<td>50</td>
+					<td>SOC</td>
+				</tr>
+				<tr>
+					<td>pSB1C3 control</td>
+					<td>1</td>
+					<td>DH5a</td>
+					<td>20</td>
+					<td>@</td>
+					<td>SOC</td>
+				</tr>
+			</table>
+</div>
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