Team:Kyoto/Notebook

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Notebook

Calendar

July
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
Octorber
1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31

アメフラシはこちら

7/15

transformation

Tatsui, Matsumoto and Sukegawa
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
BBa_J23101(Pcon)220(2/23)22LB(CP)
BBa_B0032(RBS)220(2/23)22LB(CP)

7/16

transformation

Tatsui
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
BBa_B0030(1)220(2/23)22LB(CP)
BBa_B0030(2)220(bought)22LB(CP)
BBa_B0032(1)220(2/23)22LB(CP)
BBa_B0032(2)220(bought)22LB(CP)

7/17

Colony PCR

Tatsui, Matsumoto, Li, and Yamauchi
NameⅠNameⅡ
7/16 BBa_B0030(1) Colony_17/16 BBa_B0032(1) Colony_1
7/16 BBa_B0030(1) Colony_27/16 BBa_B0032(1) Colony_2
7/16 BBa_B0030(1) Colony_37/16 BBa_B0032(1) Colony_3
7/16 BBa_B0030(2) Colony_17/16 BBa_B0032(2) Colony_1
7/16 BBa_B0030(2) Colony_27/16 BBa_B0032(2) Colony_2
7/16 BBa_B0030(2) Colony_37/16 BBa_B0032(2) Colony_3
Negative Control-
--

Liquid Culture

Tatsui, Matsumoto, Li, and Yamauchi
Namemedium
7/16 BBa_B0030(1) Colony_1Plusgrow medium(CP)
7/16 BBa_B0030(1) Colony_2Plusgrow medium(CP)
7/16 BBa_B0030(1) Colony_3Plusgrow medium(CP)
7/16 BBa_B0030(2) Colony_1Plusgrow medium(CP)
7/16 BBa_B0030(2) Colony_2Plusgrow medium(CP)
7/16 BBa_B0030(2) Colony_3Plusgrow medium(CP)
7/16 BBa_B0032(1) Colony_1Plusgrow medium(CP)
7/16 BBa_B0032(1) Colony_2Plusgrow medium(CP)
7/16 BBa_B0032(1) Colony_3Plusgrow medium(CP)
7/16 BBa_B0032(2) Colony_1Plusgrow medium(CP)
7/16 BBa_B0032(2) Colony_2Plusgrow medium(CP)
7/16 BBa_B0032(2) Colony_3Plusgrow medium(CP)

7/18

Miniprep

Tatsui, Matsumoto, Tsujii, Stephane, and Yamauchi
DNAConcentration/(µg/ml)260/280260/230
7/16 BBa_B0030(1) Colony_12291.961.40
7/16 BBa_B0030(1) Colony_22091.951.50
7/16 BBa_B0030(1) Colony_31082.122.12
7/16 BBa_B0030(2) Colony_1---
7/16 BBa_B0030(2) Colony_21742.021.56
7/16 BBa_B0030(2) Colony_31582.011.68
7/16 BBa_B0032(1) Colony_11792.002.06
7/16 BBa_B0032(1) Colony_2---
7/16 BBa_B0032(1) Colony_32211.901.37
7/16 BBa_B0032(2) Colony_12251.891.37
7/16 BBa_B0032(2) Colony_22701.981.63
7/16 BBa_B0032(2) Colony_31941.891.33

7/30

Restriction Enzyme Digestion

No name
SampleDNA/(µl)EcoR1/(µl)Xba1/(µl)Spe1/(µl)Pst1/(µl)Buffur/(µl)BSA/(µl)MilliQ/(µl)TOTAL/(µl)
7/16 BBa_B0030(2) Colony_110.30001(H)07.710
7/16 BBa_B0030(2) Colony_1100.3001(M)16.710
7/16 BBa_B0030(2) Colony_11000.301(M)07.710
7/16 BBa_B0030(2) Colony_110000.31(H)07.710
7/16 BBa_B0030(2) Colony_1(Nega)100001(M)0810

Electrophoresis

No name
LaneRestriction Enzyme Digestion Products
17/30 BBa_B0030(2) Colony_1(E CUT)
27/30 BBa_B0030(2) Colony_1(X CUT)
37/30 BBa_B0030(2) Colony_1(S CUT)
47/30 BBa_B0030(2) Colony_1(P CUT)
57/30 BBa_B0030(2) Colony_1(Nega)
61kbp ladder

7/31

Transformation

Hashitani, Tatsui, Li and Yamauchi
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
BBa_E0240(RBS-GFP-DT)22022LB(CP)
J23100(Pcon)22022LB(AMP)

Liquid Culture

No name
Namemedium
7/31 J23100(Pcon) Colony_1Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_2Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_3Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_4Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_5Plusgrow medium(AMP)
7/31 J23100(Pcon) Colony_6Plusgrow medium(AMP)

Restriction Enzyme Digestion

Hashitani, Tatsui, Li and Yamauchi
7/18 RBS SampleDNA/(µl)EcoR1/(µl)Spe1/(µl)Pst1/(µl)Xba1/(µl)Buffur/(µl)BSA/(µl)MilliQ/(µl)TOTAL/(µl)
BBa_B003250.30001(H)03.710
BBa_B0032500.3001(M)03.710
BBa_B00325000.301(H)03.710
BBa_B003250000.31(M)12.710
BBa_B0032(Nega)500001(M)0410

8/1

Electrophoresis

Honda, Li and Yamauchi
LaneRestriction Enzyme Digestion Products
11kb ladder
27/31 BBa_B0032(E CUT)
37/31 BBa_B0032(S CUT)Error
4-
5-
67/31 BBa_B0032(P CUT)
77/31 BBa_B0032(X CUT)
87/31 BBa_B0032(Nega)

PCR

Tatsui
BBa_E0240(RBS-GFP-DT)Volume
10×buffur5μL
2mMdNTPs5μL
25mM MgSO42μL
VF Primer1.5μL
VR Primer1.5μL
template0.5μL
KOD-plus1μL
MilliQ33.5μL
TOTAL50μL
StepsTemparatureTimecycle
Pre Denature94℃2min1 time
Denature94℃15sec30 cycle
Annealing55℃30sec30 cycle
Extension68℃1min30 cycle
DNAConcentration/(µg/ml)260/280260/230
Product801.911.78

8/2

Miniprep

Tatsui, Honda, Matsumoto, Yamauchi, Li, Tsujii, and Nakamura
DNAConcentration/(µg/ml)260/280260/230
7/31 J23100(Pcon) Colony_11452.113.09
7/31 J23100(Pcon) Colony_2---
7/31 J23100(Pcon) Colony_31602.013.13
7/31 J23100(Pcon) Colony_41401.872.70
7/31 J23100(Pcon) Colony_51052.395.90
7/31 J23100(Pcon) Colony_61801.901.37

8/20

Transformation

Honda
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
7/31 BBa_E0240(RBS-GFD-DT)22022LB(CP)
BBa_I73209522022LB(CP)

8/21

Colony PCR

Shimazaki
NameⅠ
8/20 BBa_I732095 Colony_1
8/20 BBa_I732095 Colony_2
8/20 BBa_I732095 Colony_3
Negative control

Electrophoresis

Honda
LanePCR Product
11kb ladder
28/21 BBa_I732095 Colony_1
38/21 BBa_I732095 Colony_2
48/21 BBa_I732095 Colony_3
5Negative Control

Liquid Culture

Honda and Tatsui
Namemedium
8/20 BBa_I732095 Colony_1Plusgrow medium(CP)
8/20 BBa_I732095 Colony_2Plusgrow medium(CP)
8/20 BBa_I732095 Colony_3Plusgrow medium(CP)

8/22

Miniprep

Honda
DNAConcentration/(µg/ml)260/280260/230
8/20 BBa_I732095 Colony_14301.682.13
8/20 BBa_I732095 Colony_22551.871.81
8/20 BBa_I732095 Colony_32151.781.35

Restriction Enzyme Digestion

Nakamura
SampleDNA/(µl)EcoR1/(µl)Spe1/(µl)Buffur/(µl)MilliQ/(µl)TOTAL/(µl)
8/22 BBa_I732095 Colony_14.7113(H)20.330
8/22 BBa_I732095 Colony_27.8113(H)17.230
8/22 BBa_I732095 Colony_39.3113(H)15.730

Transformation

Tatsui and Yamauchi
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
BBa_J04450(pSB3C5)22022LB(CP)

8/25

Transformation

Tatsui, Kita and Li
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
BBa_J04450(pSB2K3)22022LB(KAN)
BBa_C0051(pSB1A2)22022LB(AMP)

Liquid Culture

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

8/26

Miniprep

Tsujii, Honda and Matsumoto
DNAConcentration/(µg/ml)260/280260/230
8/25 BBa_J04450(pSB3C5)5751.371.37

Liquid Culture

Tatsui, Matsumoto and Nakamura
Namemedium
8/25 BBa_J04450(pSB2K3)Plusgrow medium(KAN)*5
8/25 BBa_J0051(pSB1A2)Plusgrow medium(AMP)*2

8/27

Miniprep

No Name
DNAConcentration/(µg/ml)260/280260/230
8/26 BBa_J04450(pSB2K3)10451.431.11
8/26 BBa_J0051(pSB1A2)7901.390.90

Transformation

Tatsui
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
BBa_J04450(pSB3T5)22022LB(TET)

8/28

Liquid Culture

Yamauchi
Namemedium
8/27 BBa_J04450(pSB3T5)Plusgrow medium(TET)

8/29

Miniprep

Matsumoto
DNAConcentration/(µg/ml)260/280260/230
8/28 BBa_J04450(pSB3T5)1801.601.67

Transformation

Tatsui
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
BBa_J04450(pSB4A5)22022LB(AMP)

9/1

Colony PCR

NameⅠ
-
-
-
9/1 pSB4A5 Colony_1
9/1 pSB4A5 Colony_2
Negative Control

Restriction Enzyme Digestion

No name
SampleDNA/(µl)EcoR1/(µl)Pst1/(µl)Buffur/(µl)MilliQ/(µl)TOTAL/(µl)
8/28 Miniprep pSB3T525226(H)2560
Negative control_13.7001(H)5.310
8/22 Miniprep I73209514339(H)6190
Negative control_21.5001(H)7.510
8/27 Miniprep pSB2K36339(H)6990
Negative control_31001(H)810
8/2 Miniprep Pcon Colony_3201.51.55(H)1745
Negative control_44001(H)510
8/2 Miniprep Pcon Colony_1201.51.55(H)1745
Negative control_54001(H)510

Liquid Culture

No name
Namemedium
8/29 BBa_J04450(pSB4A5) Colony_1Plusgrow medium(AMP)*2
8/1 Pcon Colony_1Plusgrow medium(AMP)*2
8/27 pSB3T5 Colony_1Plusgrow medium(TET)*2

9/2

Miniprep

Honda, Yamauchi and Li
DNAConcentration/(µg/ml)260/280260/230
8/29 BBa_J04450(pSB4A5) Colony_12101.611.18
8/1 Pcon Colony_12151.791.87
8/27 pSB3T5 Colony_12151.580.92

Gel Extractions

Matsumoto
LaneSampleVolume/(µl)
19/1 pSB3T5 Colony_1 (Nega)10
29/1 pSB3T5 Colony_1 (E+P)(Confirmation)5
39/1 pSB3T5 Colony_1 (E+P)(Extraction)55
41kb ladder3
59/1 I732095 (Nega)10
69/1 I732095 (E+P)(Confirmation)5
79/1 I732095 (E+P)(Extraction)85
81kb ladder3
99/1 pSB2K3 (Nega)10
109/1 pSB2K3 (E+P)(Confirmation)5
119/1 pSB2K3 (E+P)(Extraction)85
LaneSampleVolume/(µl)
19/1 Pcon Colony_3 (E+P)(Confirmation)5
29/1 Pcon Colony_3 (E+P)(Extraction)20
39/1 Pcon Colony_3 (E+P)(Extraction)20
49/1 Pcon Colony_3 (Nega)10
51kb ladder3
69/1 Pcon Colony_1 (E+P)(Confirmation)5
79/1 Pcon Colony_1 (E+P)(Extraction)20
89/1 Pcon Colony_1 (E+P)(Extraction)20
99/1 Pcon Colony_1 (Nega)10

9/3

Concentration Measurement

Matsumoto, Yamauchi and Li
DNAConcentration/(µg/ml)260/280260/230
pSB2K3401.480.10
Pcon Colony_1301.760.62
pSB3T5502.350.50
pSB1C3652.140.10
Pcon Colony_3451.560.58

Ligation

No name
Insert/(µL)Vector/(µL)T4 Ligase/(µl)2*Buffer/(µl)MilliQ/(µl)Total/(µl)
Pcon2.1pSB1C31.5150.410
-(Negative Control)0pSB1C31.5152.510
Pcon1.3pSB2K32.5150.210
-(Negative Control)0pSB2K32.5151.510
Pcon10pSB3T52.3150.710
-(Negative Control)0pSB3T52.3151.710

Transformation

Matsumoto
Sample/(µL)CompetentCells/(µl)Total/(µl)Medium
Pcon+pSB1C331518LB(CP)
pSB1C3(Negative Control)81018LB(CP)
Pcon+pSB2K331518LB(KAN)
pSB2K3(Negative Control)81018LB(KAN)
Pcon+pSB3T531518LB(TET)
pSB3T5(Negative Control)21012LB(TET)

9/4

Colony PCR

Honda and Yamauchi
NameⅠNameⅡNameⅢNameⅣ
-9/3 pSB1C3(Nega) Colony_79/3 Pcon+pSB3T5 Colony_59/3 Pcon+pSB2K3 Colony_1
-9/3 pSB1C3(Nega) Colony_89/3 Pcon+pSB3T5 Colony_69/3 Pcon+pSB2K3 Colony_2
9/3 pSB1C3(Nega) Colony_19/3 Pcon+pSB3T5 Colony_19/3 Pcon+pSB3T5 Colony_79/3 Pcon+pSB2K3 Colony_3
9/3 pSB1C3(Nega) Colony_29/3 Pcon+pSB3T5 Colony_29/3 Pcon+pSB3T5 Colony_89/3 Pcon+pSB2K3 Colony_4
9/3 pSB1C3(Nega) Colony_39/3 Pcon+pSB3T5 Colony_39/3 Pcon+pSB3T5 Colony_99/3 Pcon+pSB2K3 Colony_5
9/3 pSB1C3(Nega) Colony_49/3 Pcon+pSB3T5 Colony_49/3 Pcon+pSB3T5 Colony_109/3 Pcon+pSB2K3 Colony_6
9/3 pSB1C3(Nega) Colony_59/3 pSB1C3(Nega) Colony_99/3 pSB3T5(Nega) Colony_19/3 Pcon+pSB2K3 Colony_7
9/3 pSB1C3(Nega) Colony_69/3 pSB1C3(Nega) Colony_109/3 pSB3T5(Nega) Colony_29/3 Pcon+pSB2K3 Colony_8
NameⅤNameⅥNameⅦNameⅧ
9/3 Pcon+pSB2K3 Colony_99/3 Pcon+pSB1C3 Colony_29/3 Pcon+pSB1C3 Colony_109/3 pSB1C3(Nega) Colony_8
9/3 Pcon+pSB2K3 Colony_109/3 Pcon+pSB1C3 Colony_39/3 pSB1C3(Nega) Colony_19/3 pSB1C3(Nega) Colony_9
9/3 pSB2K3(Nega) Colony_19/3 Pcon+pSB1C3 Colony_49/3 pSB1C3(Nega) Colony_29/3 pSB1C3(Nega) Colony_10
9/3 pSB2K3(Nega) Colony_29/3 Pcon+pSB1C3 Colony_59/3 pSB1C3(Nega) Colony_39/3 pSB1C3(Nega) Colony_11
9/3 pSB2K3(Nega) Colony_39/3 Pcon+pSB1C3 Colony_69/3 pSB1C3(Nega) Colony_4-
9/3 pSB2K3(Nega) Colony_49/3 Pcon+pSB1C3 Colony_79/3 pSB1C3(Nega) Colony_5-
9/3 pSB2K3(Nega) Colony_59/3 Pcon+pSB1C3 Colony_89/3 pSB1C3(Nega) Colony_6-
9/3 Pcon+pSB1C3 Colony_19/3 Pcon+pSB1C3 Colony_99/3 pSB1C3(Nega) Colony_7-
NameⅨNameⅩ
9/3 Pcon+pSB1C3 Colony_118/22 Miniprep I732095 Colony_3
Negative Control-

9/5

Electrophoresis

Honda and Matsumoto
LaneColony PCR ProductsⅠLaneColony PCR ProductsⅠ
11kb ladder109/4 pSB1C3(Nega) Colony_7
2-119/4 pSB1C3(Nega) Colony_8
3-12129/4 pSB1C3(Nega) Colony_9
49/4 pSB1C3(Nega) Colony_1139/4 pSB1C3(Nega) Colony_10
59/4 pSB1C3(Nega) Colony_2149/4 Pcon+pSB2K3 Colony_1
69/4 pSB1C3(Nega) Colony_3159/4 Pcon+pSB2K3 Colony_2
79/4 pSB1C3(Nega) Colony_4169/4 Pcon+pSB2K3 Colony_3
89/4 pSB1C3(Nega) Colony_5171kb ladder
99/4 pSB1C3(Nega) Colony_6
LaneColony PCR ProductsⅡ
11kb ladder
29/4 Pcon+pSB2K3 Colony_4
39/4 Pcon+pSB2K3 Colony_5
49/4 Pcon+pSB2K3 Colony_6
59/4 Pcon+pSB2K3 Colony_7
69/4 Pcon+pSB2K3 Colony_8
79/4 Pcon+pSB2K3 Colony_9
89/4 Pcon+pSB3T5 Colony_10
99/4 pSB3T5(Nega) Colony_1
109/4 pSB3T5(Nega) Colony_2
119/4 Pcon+pSB2K3 Colony_1
129/4 Pcon+pSB2K3 Colony_2
139/4 Pcon+pSB2K3 Colony_3
149/4 Pcon+pSB2K3 Colony_4
159/4 Pcon+pSB2K3 Colony_5
169/4 Pcon+pSB2K3 Colony_6
171kb ladder
LaneColony PCR ProductsⅢ
11kb ladder
29/4 Pcon+pSB2K3 Colony_7
39/4 Pcon+pSB2K3 Colony_8
49/4 Pcon+pSB2K3 Colony_9
59/4 Pcon+pSB2K3 Colony_10
69/4 pSB2K3(Nega) Colony_1
79/4 pSB2K3(Nega) Colony_2
89/4 pSB2K3(Nega) Colony_3
99/4 pSB2K3(Nega) Colony_4
109/4 pSB2K3(Nega) Colony_5
119/4 pSB1C3(Nega) Colony_1
129/4 pSB1C3(Nega) Colony_2
139/4 pSB1C3(Nega) Colony_3
149/4 pSB1C3(Nega) Colony_4
159/4 pSB1C3(Nega) Colony_5
169/4 pSB1C3(Nega) Colony_6
171kb ladder
LaneColony PCR ProductsⅣ
11kb ladder
29/4 pSB1C3(Nega) Colony_7
39/4 pSB1C3(Nega) Colony_8
49/4 pSB1C3(Nega) Colony_9
59/4 pSB1C3(Nega) Colony_10
69/4 Pcon+pSB1C3 Colony_1
79/4 Pcon+pSB1C3 Colony_2
89/4 Pcon+pSB1C3 Colony_3
99/4 Pcon+pSB1C3 Colony_4
109/4 Pcon+pSB1C3 Colony_5
119/4 Pcon+pSB1C3 Colony_6
129/4 Pcon+pSB1C3 Colony_7
139/4 Pcon+pSB1C3 Colony_8
149/4 Pcon+pSB1C3 Colony_9
159/4 Pcon+pSB1C3 Colony_10
168/22 Miniprep BBa_I732095 Colony_3
171kb ladder
LaneColony PCR ProductsⅤ
11kb ladder
29/4 Pcon+pSB1C3 Colony_11
39/4 Negative Control
48/22 Miniprep BBa_I732095 Colony_3
5-
6-
7-
8-

9/8

9/9

9/10

9/11

9/12

9/13

9/14

9/15

9/16

9/17

9/18

9/19

9/20

9/22