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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

7/18 RBS 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)

7/31

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	30cycle
Annealing	55℃	30sec	30cycle
Extension	68℃	1min	30cycle

8/1

densitomitory after column prification

No name

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

8/2

Miniprep

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

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

Honda

	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

Shimazaki

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

8/21

Colony PCR

Shimazaki

	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

Honda

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

Honda and Tatsui

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

8/22

Miniprep

Honda

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

Nakamura

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

Tatsui and Yamauchi

BBa_J04450(pSB3C5)
DNA	2µL
Conpitent cell	20µL

8/22

Transformation

Tatsui and Yamauchi

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

8/25

Transformation

Tatsui, Kita and Li

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

Tatsui

8/22 J04450(psB3C5) 30mL

8/26

Miniprep

Tsujii, Honda and Matsumoto

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

8/26

liquid culture

Tatsui, Matsumoto and Nakamura

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

8/27

Miniprep

No name

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)

No name

DNA	6mL
Tet	60µg(10µg/mL)

8/29

Miniprep

Tatsui

DNA	Concentration(µg/ml)	260/280	260/230
7/16 BBa_B0030-(1)	229	1.96	1.40

transformation

Tatsui

Name	Sample(µl)	Competent Cells(µl)	Total(µl)	Plate
BBa_J04450	2	20(2/23)	22	LB（CP）

9/1

Colony PCR

No name

Restriction Enzyme Digestion

No name

Sample	Volume	EcoR1	Pst1	Buffur(H)	MilliQ	TOTAL
8/28 pSB3T5	25	2	2	6	25	60
Control 1	3.7	0	0	1	5.3	10
8/22 I732095	14	3	3	9	61	90
Control 2	1.5	0	0	1	7.5	10
8/27 pSB2K3	6	3	3	9	69	90
Control 3	1	0	0	1	8	10
Pcon 3	20	1.5	1.5	5	17	45
Control 4	4	0	0	1	5	10
Pcon 1	20	1.5	1.5	5	17	45
Control 5	4	0	0	1	5	10

Liquid Culture

No name

Sample	Medium	Volume
8/29 BBa_J04450(pSB4A5)	Plusgrow medium(+Amp)	12ml
8/1 Pcon 1	Plusgrow medium(+Amp)	12ml
8/27 pSB3T5 1	Plusgrow medium(+Tet)	12ml
8/29 dddD+pSB1C3 1	Plusgrow medium(+CP)	3ml
8/29 dddD+pSB1C3 2	Plusgrow medium(+CP)	3ml
8/29 dddD+pSB1C3 3	Plusgrow medium(+CP)	3ml

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

7/31

Restriction Enzyme Digestion

8/1

Electrophoresis for confirmation

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)

8/29

Miniprep

transformation

9/1

Colony PCR

Restriction Enzyme Digestion

Liquid Culture