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Notebook

You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very ustoeful organizational tool as well.

7/15

transformation

Name	Sample(µl)	Competent Cells(µl)	Total(µl)	Plate
BBa_J23101	2	20(2/23)	22	LB（CP）
BBA_B0032	2	20（2/23）	22	LB（CP）

7/16

transformation

Name	Sample(µl)	Competent Cells(µl)	Total(µl)	Plate
BBa_B0030	2	20(2/23)	22	LB（CP）
BBA_B0032	2	20（2/23）	22	LB（CP）

7/17

Colony PCR

Sample
7/16 BBa_B0030-(1)
7/16 BBa_B0030-(2)
7/16 BBa_B0030-(3)
7/16 BBa_B0030-(4)
7/16 BBa_B0030-(5)
7/16 BBa_B0030-(6)
7/16 BBa_B0032-(1)
7/16 BBa_B0032-(2)
7/16 BBa_B0032-(3)
7/16 BBa_B0032-(4)
7/16 BBa_B0032-(5)
7/16 BBa_B0032-(6)
NC

Liquid Culture

Sample	medium
7/16 BBa_B0030-(1)	Plusgrow medium(+CP)
7/16 BBa_B0030-(2)	Plusgrow medium(+CP)
7/16 BBa_B0030-(3)	Plusgrow medium(+CP)
7/16 BBa_B0030-(4)	Plusgrow medium(+CP)
7/16 BBa_B0030-(5)	Plusgrow medium(+CP)
7/16 BBa_B0030-(6)	Plusgrow medium(+CP)
7/16 BBa_B0032-(1)	Plusgrow medium(+CP)
7/16 BBa_B0032-(2)	Plusgrow medium(+CP)
7/16 BBa_B0032-(3)	Plusgrow medium(+CP)
7/16 BBa_B0032-(4)	Plusgrow medium(+CP)
7/16 BBa_B0032-(5)	Plusgrow medium(+CP)
7/16 BBa_B0032-(6)	Plusgrow medium(+CP)

7/18

Miniprep

DNA	Concentration(µg/ml)	260/280	260/230
7/16 BBa_B0030-(1)	229	1.96	1.40
7/16 BBa_B0030-(2)	209	1.95	1.50
7/16 BBa_B0030-(3)	108	2.12	2.12
7/16 BBa_B0030-(4)	186	-	-
7/16 BBa_B0030-(5)	174	2.02	1.56
7/16 BBa_B0030-(6)	158	2.01	1.68
7/16 BBa_B0032-(1)	179	2.00	2.06
7/16 BBa_B0032-(3)	221	1.90	1.37
7/16 BBa_B0032-(4)	225	1.89	1.37
7/16 BBa_B0032-(5)	270	1.98	1.63
7/16 BBa_B0032-(6)	194	1.89	1.33

7/28

Restriction Enzyme Digestion

7/18 RBS Sample	EcoR1	Xba1	Spe1	Pst1	Buffur	BSA	MilliQ	TOTAL
Sample1	0.3	0	0	0	1(H)	0	7.7	10
Sample2	0	0.3	0	0	1(M)	1	6.7	10
Sample3	0	0	0.3	0	1(M)	0	7.7	10
Sample4	0	0	0	0.3	1(H)	0	7.7	10
Control	0	0	0	0	1	0	8	10

7/30

Restriction Enzyme Digestion

7/18 RBS Sample	EcoR1	Xba1	Spe1	Pst1	Buffur	BSA	MilliQ	TOTAL
Sample1	0.3	0	0	0	1(H)	0	7.7	10
Sample2	0	0.3	0	0	1(M)	1	6.7	10
Sample3	0	0	0.3	0	1(M)	0	7.7	10
Sample4	0	0	0	0.3	1(H)	0	7.7	10
Control	0	0	0	0	1	0	8	10

7/30

Electrophoresis for confirmation

Lane	Restriction Enzyme Digestion Products
1	Sample1
2	Sample2
3	Sample3
4	Sample4
5	Sample5
6	1kbp ladder

7/31

Transformation

	2-24B BBa_E0240(RBS-GFP-DT)+CP	4-17D J23100(Pcon) +Amp
plasmid	2μL	2μL
DH5α	20μL	2μL
TOTAL	22μL	22μL

7/31

Restriction Enzyme Digestion

Sample	Plasmid	EcoR1	Xba1	Spe1	Pst1	Buffur	BSA	MilliQ	TOTAL
Sample1	5	0.3	0	0	0	1(H)	0	3.7	10
Sample2	5	0	0.3	0	0	1(M)	0	3.7	10
Sample3	5	0	0	0.3	0	1(H)	0	3.7	10
Sample4	5	0	0	0	0.3	1(M)	1	2.7	10
Control	5	0	0	0	0	1(M)	0	4	10

8/1

Electrophoresis for confirmation

Lane	Sample
1	1kb ladder
2	7/31 Restruction Enzyme Digestion Products Sample1
3	7/31 Restruction Enzyme Digestion Products Sample2 Error
4	NC
5	NC
6	7/31 Restruction Enzyme Digestion Products Sample3
7	7/31 Restruction Enzyme Digestion Products Sample4
8	7/31 Restruction Enzyme Digestion Products Sample5

8/1

PCR

BBa_E0240(RBS-GFP-DT)
	Volume
10×buffur	5μL
2mMdNTPs	5μL
25mM MgSO4	2μL
VF Primer	1.5μL
VR Primer	1.5μL
template	0.5μL
KOD-plus	1μL
MilliQ	33.5μL
TOTAL	50μL

8/1

PCR

BBa_E0240(RBS-GFP-DT)
	Temparature	Time
Pre Denature	94℃	2min
Denature	94℃	15sec	30cycle
Anniling	55℃	30sec	30cycle
Exensim	68℃	1min	30cycle

8/1

densitomitory after column prification

BBa_E0240(RBS-GFP-DT)
Density	260/230	260/280
80μg/mL	1.78	1.91

8/2

Miniprep

7/31 Pcon
Sample	Density(μg/mL)	260/280	260/230
1	2.9	2.11	3.09
2	1.3	2.59	-
3	3.2	2.01	3.13
4	2.8	1.87	2.70
5	2.1	2.39	5.90
6	3.6	1.90	1.37

8/20

Transformation

	BBa_I732095	7/31 Transformation BBa_E0240(RBS-aFD-DT)(+CP)
plasmid	2μL	2μL
DH5α	20μL	20μL
TOTAL	22μL	22μL
	×2	×2

8/21

Colony PCR

	Master Mix
Quick Taq	20μL
VF	0.8μL
VR	0.8μL
MilliQ	18.4μL
TOTAL	40μL

8/21

Colony PCR

	Temparature	Time
Pre Denature	94.0℃	2min
Denature	94.0℃	30sec
Anniling	55.0℃	0.8μL
Extention	68.0℃	18.4μL
Denature - Extention 30 cycle

8/21

Electrophoresis for confirmation

Lane	Sample
1	1kb ladder
2	BBa_E0430-Sample1
3	BBa_E0430-Sample2
4	BBa_E0430-Sample3
5	Negative Control

8/21

Electrophoresis for confirmation

1	BBa_E0430-Sample1
2	BBa_E0430-Sample2
3	BBa_E0430-Sample3

8/22

Miniprep

Sample	density	260/280	260/230
BBa_I732095-Sample1	430µg/mL	1.68	2.13
BBa_I732095-Sample2	255µg/mL	1.87	1.81
BBa_I732095-Sample3	215µg/mL	1.78	1.35

8/22

Restriction Enzyme Digestion

BBa_I732095-Sample1	4.7µL
EcoRI	1µL
SpeRI	1µL
Buffur(H)	3µL
MilliQ	20.3µL
TOTAL	30µL
BBa_I732095-Sample2	7.8µL
EcoRI	1µL
SpeRI	1µL
Buffur(H)	3µL
MilliQ	20.3µL
TOTAL	30µL
BBa_I732095-Sample3	9.3µL
EcoRI	1µL
SpeRI	1µL
Buffur(H)	3µL
MilliQ	15.7µL
TOTAL	30µL

8/22

Transformation

BBa_J04450(pSB3C5)
DNA	2µL
Conpitent cell	20µL

8/22

Transformation

BBa_J04450(pSB3C5)	Time
on ice	30min
heat shock	45sec
on ice	2min
Preculture	1hour

8/25

Transformation

BBa_J04450(2014)
DNA	20µL
Conpitent cell	40µL
BBa_J04450(2013)
DNA	20µL
Conpitent cell	40µL
BBa_C0051
DNA	20µL
Conpitent cell	40µL

8/25

liquid culture

8/22 J04450(psB3C5) 30mL

8/26

Miniprep

Sample	density(µg/mL)	260/280	260/230
J04450(psB3C5)	575	1.37	1.34

8/26

liquid culture

8/25 psB2K3×5(30mL)
8/25 psB1A2×2(12mL)

8/27

Miniprep

Sample	density	260/280	260/230
8/26 licuid culture psB2K3	1045(µg/mL)	1.43	1.11
8/26 licuid culture psB1A2	790(µg/mL)	1.39	0.90

8/27

BBa_J04450(psB3T5)

DNA	6mL
Tet	60µg(10µg/mL)

@@ Line 205: / Line 205: @@
 </table>
+<h2>7/30</h2>
+<h3>Restriction Enzyme Digestion</h3>
+<table class="note">
+<tr><th>7/18 RBS Sample</th><th>EcoR1</th><th>Xba1</th><th>Spe1</th><th>Pst1</th><th>Buffur</th><th>BSA</th><th>MilliQ</th><th>TOTAL</tr>
+<tr><td>Sample1</td><td>0.3</td><td>0</td><td>0</td><td>0</td><td>1(H)</td><td>0</td><td>7.7</td><td>10</td></tr>
+<tr><td>Sample2</td><td>0</td><td>0.3</td><td>0</td><td>0</td><td>1(M)</td><td>1</td><td>6.7</td><td>10</td></tr>
+<tr><td>Sample3</td><td>0</td><td>0</td><td>0.3</td><td>0</td><td>1(M)</td><td>0</td><td>7.7</td><td>10</td></tr>
+<tr><td>Sample4</td><td>0</td><td>0</td><td>0</td><td>0.3</td><td>1(H)</td><td>0</td><td>7.7</td><td>10</td></tr>
+<tr><td>Control</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>0</td><td>8</td><td>10</td></tr>
+</table>
+<h2>7/30</h2>
+<h3>Electrophoresis for confirmation</h3>
+<table class="note">
+<tr><th>Lane</th><th>Restriction Enzyme Digestion Products</td></tr>
+<tr><td>1</td><td>Sample1</td></tr>
+<tr><td>2</td><td>Sample2</td></tr>
+<tr><td>3</td><td>Sample3</td></tr>
+<tr><td>4</td><td>Sample4</td></tr>
+<tr><td>5</td><td>Sample5</td></tr>
+<tr><td>6</td><td>1kbp ladder</td></tr>
+</table>
+<h2>7/31</h2>
+<h3>Transformation</h3>
+<table class="note">
+<tr><th></th><th>2-24B BBa_E0240(RBS-GFP-DT)+CP</th><th>4-17D J23100(Pcon) +Amp</th></tr>
+<tr><th>plasmid</th><th>2μL</th><th>2μL</th></tr>
+<tr><th>DH5α</th><th>20μL</th><th>2μL</th></tr>
+<tr><th>TOTAL</th><th>22μL</th><th>22μL</th></tr>
+</table>
+<h2>7/31</h2>
+<h3>Restriction Enzyme Digestion</h3>
+<table class="note">
+<tr><th>Sample</th><th>Plasmid</th><th>EcoR1</th><th>Xba1</th><th>Spe1</th><th>Pst1</th><th>Buffur</th><th>BSA</th><th>MilliQ</th><th>TOTAL</tr>
+<tr><td>Sample1</td><td>5</td><td>0.3</td><td>0</td><td>0</td><td>0</td><td>1(H)</td><td>0</td><td>3.7</td><td>10</td></tr>
+<tr><td>Sample2</td><td>5</td><td>0</td><td>0.3</td><td>0</td><td>0</td><td>1(M)</td><td>0</td><td>3.7</td><td>10</td></tr>
+<tr><td>Sample3</td><td>5</td><td>0</td><td>0</td><td>0.3</td><td>0</td><td>1(H)</td><td>0</td><td>3.7</td><td>10</td></tr>
+<tr><td>Sample4</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0.3</td><td>1(M)</td><td>1</td><td>2.7</td><td>10</td></tr>
+<tr><td>Control</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(M)</td><td>0</td><td>4</td><td>10</td></tr>
+</table>
+<h2>8/1</h2>
+<h3>Electrophoresis for confirmation</h3>
+<table class="note">
+<tr><th>Lane</th><th>Sample</th></tr>
+<tr><th>1</th><th>1kb ladder</th></tr>
+<tr><th>2</th><th>7/31 Restruction Enzyme Digestion Products Sample1</th></tr>
+<tr><th>3</th><th>7/31 Restruction Enzyme Digestion Products Sample2 Error</th></tr>
+<tr><th>4</th><th>NC</th></tr>
+<tr><th>5</th><th>NC</th></tr>
+<tr><th>6</th><th>7/31 Restruction Enzyme Digestion Products Sample3</th></tr>
+<tr><th>7</th><th>7/31 Restruction Enzyme Digestion Products Sample4</th></tr>
+<tr><th>8</th><th>7/31 Restruction Enzyme Digestion Products Sample5</th></tr>
+</table>
+<h2>8/1</h2>
+<h3>PCR</h3>
+<table class="note">
+<tr><th>BBa_E0240(RBS-GFP-DT)</th></tr>
+<tr><th></th><th>Volume</th></tr>
+<tr><th>10×buffur</th><th>5μL</th></tr>
+<tr><th>2mMdNTPs</th><th>5μL</th></tr>
+<tr><th>25mM MgSO4</th><th>2μL</th></tr>
+<tr><th>VF Primer</th><th>1.5μL</th></tr>
+<tr><th>VR Primer</th><th>1.5μL</th></tr>
+<tr><th>template</th><th>0.5μL</th></tr>
+<tr><th>KOD-plus</th><th>1μL</th></tr>
+<tr><th>MilliQ</th><th>33.5μL</th></tr>
+<tr><th>TOTAL</th><th>50μL</th></tr>
+</table>
+<h2>8/1</h2>
+<h3>PCR</h3>
+<table class="note">
+<tr><th>BBa_E0240(RBS-GFP-DT)</th></tr>
+<tr><td></td><td>Temparature</td><td>Time</td></tr>
+<tr><td>Pre Denature</td><td>94℃</td><td>2min</td></tr>
+<tr><td>Denature</td><td>94℃</td><td>15sec</td><td>30cycle</td></tr>
+<tr><td>Anniling</td><td>55℃</td><td>30sec</td><td>30cycle</td></tr>
+<tr><td>Exensim</td><td>68℃</td><td>1min</td><td>30cycle</td></tr>
+</table>
+<h2>8/1</h2>
+<h3>densitomitory after column prification</h3>
+<table class="note">
+<tr><th>BBa_E0240(RBS-GFP-DT)</th></tr>
+<tr><td>Density</td><td>260/230</td><td>260/280</td></tr>
+<tr><td>80μg/mL</td><td>1.78</td><td>1.91</td></tr>
+</table>
+<h2>8/2</h2>
+<h3>Miniprep</h3>
+<table class="note">
+<tr><th>7/31 Pcon</th></tr>
+<tr><td>Sample</td><td>Density(μg/mL)</td><td>260/280</td><td>260/230</td></tr>
+<tr><td>1</td><td>2.9</td><td>2.11</td><td>3.09</td></tr>
+<tr><td>2</td><td>1.3</td><td>2.59</td><td>-</td></tr>
+<tr><td>3</td><td>3.2</td><td>2.01</td><td>3.13</td></tr>
+<tr><td>4</td><td>2.8</td><td>1.87</td><td>2.70</td></tr>
+<tr><td>5</td><td>2.1</td><td>2.39</td><td>5.90</td></tr>
+<tr><td>6</td><td>3.6</td><td>1.90</td><td>1.37</td></tr>
+</table>
+<h2>8/20</h2>
+<h3>Transformation</h3>
+<table class="note">
+<tr><th></th><th>BBa_I732095</th><th>7/31 Transformation BBa_E0240(RBS-aFD-DT)(+CP)</th></tr>
+<tr><td>plasmid</td><td>2μL</td><td>2μL</td></tr>
+<tr><td>DH5α</td><td>20μL</td><td>20μL</td></tr>
+<tr><td>TOTAL</td><td>22μL</td><td>22μL</td></tr>
+<tr><td></td><td>×2</td><td>×2</td></tr>
+</table>
+<h2>8/21</h2>
+<h3>Colony PCR</h3>
+<table class="note">
+<tr><th></th><th>Master Mix</th></tr>
+<tr><td>Quick Taq</td><td>20μL</td></tr>
+<tr><td>VF</td><td>0.8μL</td></tr>
+<tr><td>VR</td><td>0.8μL</td></tr>
+<tr><td>MilliQ</td><td>18.4μL</td></tr>
+<tr><td>TOTAL</td><td>40μL</td></tr>
+</table>
+<h2>8/21</h2>
+<h3>Colony PCR</h3>
+<table class="note">
+<tr><th></th><th>Temparature</th><th>Time</th></tr>
+<tr><td>Pre Denature</td><td>94.0℃</td><td>2min</td></tr>
+<tr><td>Denature</td><td>94.0℃</td><td>30sec</td></tr>
+<tr><td>Anniling</td><td>55.0℃</td><td>0.8μL</td></tr>
+<tr><td>Extention</td><td>68.0℃</td><td>18.4μL</td></tr>
+<tr><td>Denature - Extention 30 cycle</td></tr>
+</table>
+<h2>8/21</h2>
+<h3>Electrophoresis for confirmation</h3>
+<table class="note">
+<tr><th>Lane</th><th>Sample</th></tr>
+<tr><td>1</td><td>1kb ladder</td></tr>
+<tr><td>2</td><td>BBa_E0430-Sample1</td></tr>
+<tr><td>3</td><td>BBa_E0430-Sample2</td></tr>
+<tr><td>4</td><td>BBa_E0430-Sample3</td></tr>
+<tr><td>5</td><td>Negative Control</td></tr>
+</table>
+<h2>8/21</h2>
+<h3>Electrophoresis for confirmation</h3>
+<table class="note">
+<tr><th>1</th><th>BBa_E0430-Sample1</th></tr>
+<tr><td>2</td><td>BBa_E0430-Sample2</td></tr>
+<tr><td>3</td><td>BBa_E0430-Sample3</td></tr>
+</table>
+<h2>8/22</h2>
+<h3>Miniprep</h3>
+<table class="note">
+<tr><th>Sample</th><th>density</th><th>260/280</th><th>260/230</th></tr>
+<tr><td>BBa_I732095-Sample1</td><td>430µg/mL</td><td>1.68</td><td>2.13</td></tr>
+<tr><td>BBa_I732095-Sample2</td><td>255µg/mL</td><td>1.87</td><td>1.81</td></tr>
+<tr><td>BBa_I732095-Sample3</td><td>215µg/mL</td><td>1.78</td><td>1.35</td></tr>
+</table>
+<h2>8/22</h2>
+<h3>Restriction Enzyme Digestion</h3>
+<table class="note">
+<tr><td>BBa_I732095-Sample1</td><td>4.7µL</td></tr>
+<tr><td>EcoRI</td><td>1µL</td></tr>
+<tr><td>SpeRI</td><td>1µL</td></tr>
+<tr><td>Buffur(H)</td><td>3µL</td></tr>
+<tr><td>MilliQ</td><td>20.3µL</td></tr>
+<tr><td>TOTAL</td><td>30µL</td></tr>
+<tr><td>BBa_I732095-Sample2</td><td>7.8µL</td></tr>
+<tr><td>EcoRI</td><td>1µL</td></tr>
+<tr><td>SpeRI</td><td>1µL</td></tr>
+<tr><td>Buffur(H)</td><td>3µL</td></tr>
+<tr><td>MilliQ</td><td>20.3µL</td></tr>
+<tr><td>TOTAL</td><td>30µL</td></tr>
+<tr><td>BBa_I732095-Sample3</td><td>9.3µL</td></tr>
+<tr><td>EcoRI</td><td>1µL</td></tr>
+<tr><td>SpeRI</td><td>1µL</td></tr>
+<tr><td>Buffur(H)</td><td>3µL</td></tr>
+<tr><td>MilliQ</td><td>15.7µL</td></tr>
+<tr><td>TOTAL</td><td>30µL</td></tr>
+</table>
+<h2>8/22</h2>
+<h3>Transformation</h3>
+<table class="note">
+<tr><th>BBa_J04450(pSB3C5)</th></tr>
+<tr><td>DNA</td><td>2µL</td></tr>
+<tr><td>Conpitent cell</td><td>20µL</td></tr>
+</table>
+<h2>8/22</h2>
+<h3>Transformation</h3>
+<table class="note">
+<tr><th>BBa_J04450(pSB3C5)</th><th>Time</th></tr>
+<tr><td>on ice</td><td>30min</td></tr>
+<tr><td>heat shock</td><td>45sec</td></tr>
+<tr><td>on ice</td><td>2min</td></tr>
+<tr><td>Preculture</td><td>1hour</td></tr>
+</table>
+<h2>8/25</h2>
+<h3>Transformation</h3>
+<table class="note">
+<tr><th>BBa_J04450(2014)</th></tr>
+<tr><td>DNA</td><td>20µL</td></tr>
+<tr><td>Conpitent cell</td><td>40µL</td></tr>
+<tr><th>BBa_J04450(2013)</th></tr>
+<tr><td>DNA</td><td>20µL</td></tr>
+<tr><td>Conpitent cell</td><td>40µL</td></tr>
+<tr><th>BBa_C0051</th></tr>
+<tr><td>DNA</td><td>20µL</td></tr>
+<tr><td>Conpitent cell</td><td>40µL</td></tr>
+</table>
+<h2>8/25</h2>
+<h3>liquid culture</h3>
+<table class="note">
+<tr><th>8/22 J04450(psB3C5) 30mL</th></tr>
+</table>
+<h2>8/26</h2>
+<h3>Miniprep</h3>
+<table class="note">
+<tr><th>Sample</th><th>density(µg/mL)</td><td>260/280</td><td>260/230</th></tr>
+<tr><td>J04450(psB3C5)</td><td>575</td><td>1.37</td><td>1.34</td></tr>
+</table>
+<h2>8/26</h2>
+<h3>liquid culture</h3>
+<table class="note">
+<tr><th>8/25 psB2K3×5(30mL)</th></tr>
+<tr><th>8/25 psB1A2×2(12mL)</th></tr>
+</table>
+<h2>8/27</h2>
+<h3>Miniprep</h3>
+<table class="note">
+<tr><th>Sample</th><th>density</th><th>260/280</th><th>260/230</th></tr>
+<tr><td>8/26 licuid culture psB2K3</td><td>1045(µg/mL)</td><td>1.43</td><td>1.11</td></tr>
+<tr><td>8/26 licuid culture psB1A2</td><td>790(µg/mL)</td><td>1.39</td><td>0.90</td></tr>
+</table>
+<h2>8/27</h2>
+<h3>BBa_J04450(psB3T5)</h3>
+<table class="note">
+<tr><th>DNA</th><th>6mL</th></tr>
+<tr><th>Tet</th><th>60µg(10µg/mL)</th></tr>
+</table>

Team:Kyoto/Notebook

From 2014.igem.org

Revision as of 02:47, 29 August 2014

Notebook

7/15

transformation

7/16

transformation

7/17

Colony PCR

Liquid Culture

7/18

Miniprep

7/28

Restriction Enzyme Digestion

7/30

Restriction Enzyme Digestion

7/30

Electrophoresis for confirmation

7/31

Transformation

7/31

Restriction Enzyme Digestion

8/1

Electrophoresis for confirmation

8/1

PCR

8/1

PCR

8/1

densitomitory after column prification

8/2

Miniprep

8/20

Transformation

8/21

Colony PCR

8/21

Colony PCR

8/21

Electrophoresis for confirmation

8/21

Electrophoresis for confirmation

8/22

Miniprep

8/22

Restriction Enzyme Digestion

8/22

Transformation

8/22

Transformation

8/25

Transformation

8/25

liquid culture

8/26

Miniprep

8/26

liquid culture

8/27

Miniprep

8/27

BBa_J04450(psB3T5)