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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.

アメフラシはこちら

7/15

transformation

Tatsui, Matsumoto and Sukegawa

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Medium
BBa_J23101(Pcon)	2	20(2/23)	22	LB(CP)
BBa_B0032(RBS)	2	20(2/23)	22	LB(CP)

7/16

transformation

Tatsui

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Medium
BBa_B0030(1)	2	20(2/23)	22	LB(CP)
BBa_B0030(2)	2	20(bought)	22	LB(CP)
BBa_B0032(1)	2	20(2/23)	22	LB(CP)
BBa_B0032(2)	2	20(bought)	22	LB(CP)

7/17

Colony PCR

Tatsui, Matsumoto, Li, and Yamauchi

Name
7/16 BBa_B0030(1) Colony_1
7/16 BBa_B0030(1) Colony_2
7/16 BBa_B0030(1) Colony_3
7/16 BBa_B0030(2) Colony_1
7/16 BBa_B0030(2) Colony_2
7/16 BBa_B0030(2) Colony_3
7/16 BBa_B0032(1) Colony_1
7/16 BBa_B0032(1) Colony_2
7/16 BBa_B0032(1) Colony_3
7/16 BBa_B0032(2) Colony_1
7/16 BBa_B0032(2) Colony_2
7/16 BBa_B0032(2) Colony_3
Negative control

Liquid Culture

Tatsui, Matsumoto, Li, and Yamauchi

Name	medium
7/16 BBa_B0030(1) Colony_1	Plusgrow medium(CP)
7/16 BBa_B0030(1) Colony_2	Plusgrow medium(CP)
7/16 BBa_B0030(1) Colony_3	Plusgrow medium(CP)
7/16 BBa_B0030(2) Colony_1	Plusgrow medium(CP)
7/16 BBa_B0030(2) Colony_2	Plusgrow medium(CP)
7/16 BBa_B0030(2) Colony_3	Plusgrow medium(CP)
7/16 BBa_B0032(1) Colony_1	Plusgrow medium(CP)
7/16 BBa_B0032(1) Colony_2	Plusgrow medium(CP)
7/16 BBa_B0032(1) Colony_3	Plusgrow medium(CP)
7/16 BBa_B0032(2) Colony_1	Plusgrow medium(CP)
7/16 BBa_B0032(2) Colony_2	Plusgrow medium(CP)
7/16 BBa_B0032(2) Colony_3	Plusgrow medium(CP)

7/18

Miniprep

Tatsui, Matsumoto, Tsujii, Stephane, and Yamauchi

DNA	Concentration/(µg/ml)	260/280	260/230
7/16 BBa_B0030(1) Colony_1	229	1.96	1.40
7/16 BBa_B0030(1) Colony_2	209	1.95	1.50
7/16 BBa_B0030(1) Colony_3	108	2.12	2.12
7/16 BBa_B0030(2) Colony_1	-	-	-
7/16 BBa_B0030(2) Colony_2	174	2.02	1.56
7/16 BBa_B0030(2) Colony_3	158	2.01	1.68
7/16 BBa_B0032(1) Colony_1	179	2.00	2.06
7/16 BBa_B0032(1) Colony_2	-	-	-
7/16 BBa_B0032(1) Colony_3	221	1.90	1.37
7/16 BBa_B0032(2) Colony_1	225	1.89	1.37
7/16 BBa_B0032(2) Colony_2	270	1.98	1.63
7/16 BBa_B0032(2) Colony_3	194	1.89	1.33

7/30

Restriction Enzyme Digestion

No name

Sample	Sample/(µl)	EcoR1/(µl)	Xba1/(µl)	Spe1/(µl)	Pst1/(µl)	Buffur/(µl)	BSA/(µl)	MilliQ/(µl)	TOTAL/(µl)
7/16 BBa_B0030(2) Colony_1	1	0.3	0	0	0	1(H)	0	7.7	10
7/16 BBa_B0030(2) Colony_1	1	0	0.3	0	0	1(M)	1	6.7	10
7/16 BBa_B0030(2) Colony_1	1	0	0	0.3	0	1(M)	0	7.7	10
7/16 BBa_B0030(2) Colony_1	1	0	0	0	0.3	1(H)	0	7.7	10
Control	1	0	0	0	0	1(M)	0	8	10

Electrophoresis for confirmation

No name

Lane	Restriction Enzyme Digestion Products
1	Product1
2	Product2
3	Product3
4	Product4
5	Product5
6	1kbp ladder

7/31

Transformation

Hashitani, Tatsui, Li and Yamauchi

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Medium
BBa_E0240(RBS-GFP-DT)	2	20	22	LB(CP)
J23100(Pcon)	2	20	22	LB(AMP)

Liquid Culture

No name

Name	medium
7/31 J23100(Pcon) Colony_1	Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_2	Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_3	Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_4	Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_5	Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_6	Plusgrow medium(AMP)

Restriction Enzyme Digestion

Hashitani, Tatsui, Li and Yamauchi

7/18 RBS Sample	Sample/(µl)	EcoR1/(µl)	Spe1/(µl)	Pst1/(µl)	Xba1/(µl)	Buffur/(µl)	BSA/(µl)	MilliQ/(µl)	TOTAL/(µl)
BBa_B0032	5	0.3	0	0	0	1(H)	0	3.7	10
BBa_B0032	5	0	0.3	0	0	1(M)	0	3.7	10
BBa_B0032	5	0	0	0.3	0	1(H)	0	3.7	10
BBa_B0032	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

Honda, Li and Yamauchi

Lane	Restriction Enzyme Digestion Products
1	1kb ladder
2	Product1
3	Product2 Error
4	-
5	-
6	Product3
7	Product4
8	Product5

8/1

PCR

Tatsui

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

Steps	Temparature	Time	cycle
Pre Denature	94℃	2min	1 time
Denature	94℃	15sec	30 cycle
Annealing	55℃	30sec	30 cycle
Extension	68℃	1min	30 cycle

DNA	Concentration/(µg/ml)	260/280	260/230
Product	80	1.91	1.78

8/2

Miniprep

Tatsui, Honda, Matsumoto, Yamauchi, Li, Tsujii, and Nakamura

DNA	Concentration/(µg/ml)	260/280	260/230
7/31 J23100(Pcon) Colony_1	145	2.11	3.09
7/31 J23100(Pcon) Colony_2	-	-	-
7/31 J23100(Pcon) Colony_3	160	2.01	3.13
7/31 J23100(Pcon) Colony_4	140	1.87	2.70
7/31 J23100(Pcon) Colony_5	105	2.39	5.90
7/31 J23100(Pcon) Colony_6	180	1.90	1.37

8/20

Transformation

Honda

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Medium
7/31 BBa_E0240(RBS-GFD-DT)	2	20	22	LB(CP)
BBa_I732095	2	20	22	LB(CP)

8/21

Colony PCR

Shimazaki

Name
8/20 BBa_I732095 Colony_1
8/20 BBa_I732095 Colony_2
8/20 BBa_I732095 Colony_3
Negative control

Electrophoresis for confirmation

Honda

Lane	PCR Product
1	1kb ladder
2	8/21 BBa_I732095 Colony_1
3	8/21 BBa_I732095 Colony_2
4	8/21 BBa_I732095 Colony_3
5	Negative Control

Liquid Culture

Honda and Tatsui

Name	medium
8/20 BBa_I732095 Colony_1	Plusgrow medium(CP)
8/20 BBa_I732095 Colony_2	Plusgrow medium(CP)
8/20 BBa_I732095 Colony_3	Plusgrow medium(CP)

8/22

Miniprep

Honda

DNA	Concentration/(µg/ml)	260/280	260/230
8/20 BBa_I732095 Colony_1	430	1.68	2.13
8/20 BBa_I732095 Colony_2	255	1.87	1.81
8/20 BBa_I732095 Colony_3	215	1.78	1.35

Restriction Enzyme Digestion

Nakamura

Sample	Sample/(µl)	EcoR1/(µl)	Spe1/(µl)	Buffur/(µl)	MilliQ/(µl)	TOTAL/(µl)
8/22 BBa_I732095 Colony_1	4.7	1	1	3(H)	20.3	30
8/22 BBa_I732095 Colony_2	7.8	1	1	3(H)	17.2	30
8/22 BBa_I732095 Colony_3	9.3	1	1	3(H)	15.7	30

Transformation

Tatsui and Yamauchi

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Medium
BBa_J04450(pSB3C5)	2	20	22	LB(CP)

8/25

Transformation

Tatsui, Kita and Li

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Medium
BBa_J04450(pSB2K3)	2	20	22	LB(KAN)
BBa_C0051(pSB1A2)	2	20	22	LB(AMP)

Liquid Culture

Tatsui

Name	medium
8/22 BBa_J04450(pSB3C5)	Plusgrow medium(CP)*5

8/26

Miniprep

Tsujii, Honda and Matsumoto

DNA	Concentration/(µg/ml)	260/280	260/230
8/25 BBa_J04450(pSB3C5)	575	1.37	1.37

Liquid Culture

Tatsui, Matsumoto and Nakamura

Name	medium
8/25 BBa_J04450(pSB2K3)	Plusgrow medium(KAN)*5
8/25 BBa_J0051(pSB1A2)	Plusgrow medium(AMP)*2

8/27

Miniprep

No Name

DNA	Concentration/(µg/ml)	260/280	260/230
8/26 BBa_J04450(pSB2K3)	1045	1.43	1.11
8/26 BBa_J0051(pSB1A2)	790	1.39	0.90

Transformation

Tatsui

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Medium
BBa_J04450(pSB3T5)	2	20	22	LB(TET)

8/28

Liquid Culture

Yamauchi

Name	medium
8/27 BBa_J04450(pSB3T5)	Plusgrow medium(TET)

8/29

Miniprep

Matsumoto

DNA	Concentration/(µg/ml)	260/280	260/230
8/28 BBa_J04450(pSB3T5)	180	1.60	1.67

Transformation

Tatsui

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Medium
BBa_J04450(pSB4A5)	2	20	22	LB(AMP)

9/1

Colony PCR

あめふらし

Restriction Enzyme Digestion

No name

Sample	Sample/(µl)	EcoR1/(µl)	Pst1/(µl)	Buffur/(µl)	MilliQ/(µl)	TOTAL/(µl)
8/28 Miniprep pSB3T5	25	2	2	6(H)	25	60
Negative control_1	3.7	0	0	1(H)	5.3	10
8/22 Miniprep I732095	14	3	3	9(H)	61	90
Negative control_2	1.5	0	0	1(H)	7.5	10
8/27 Miniprep pSB2K3	6/td>	3	3	9(H)	69	90
Negative control_3	1	0	0	1(H)	8	10
8/2 Miniprep Pcon Colony_3	20	1.5	1.5	5(H)	17	45
Negative control_4	4	0	0	1(H)	5	10
8/2 Miniprep Pcon Colony_1	20	1.5	1.5	5(H)	17	45
Negative control_5	4	0	0	1(H)	5	10

Liquid Culture

No name

Name	medium
8/29 BBa_J04450(pSB4A5) Colony_1	Plusgrow medium(AMP)*2
8/1 Pcon Colony_1	Plusgrow medium(AMP)*2
8/27 pSB3T5 Colony_1	Plusgrow medium(TET)*2

Team:Kyoto/Notebook

From 2014.igem.org

Notebook

7/15

transformation

7/16

transformation

7/17

Colony PCR

Liquid Culture

7/18

Miniprep

7/30

Restriction Enzyme Digestion

Electrophoresis for confirmation

7/31

Transformation

Liquid Culture

Restriction Enzyme Digestion

8/1

Electrophoresis for confirmation

8/1

PCR

8/2

Miniprep

8/20

Transformation

8/21

Colony PCR

Electrophoresis for confirmation

Liquid Culture

8/22

Miniprep

Restriction Enzyme Digestion

Transformation

8/25

Transformation

Liquid Culture

8/26

Miniprep

Liquid Culture

8/27

Miniprep

Transformation

8/28

Liquid Culture

8/29

Miniprep

Transformation

9/1

Colony PCR

Restriction Enzyme Digestion

Liquid Culture