Team:KIT-Kyoto/Notebook/Labnote
From 2014.igem.org
LabNote
June
June 10, 2014
OKAMOTO
Miniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3) | pGEX6P-2 | CuMTSE1 |
BL21(DE3) | pGEX6P-2 | CuMTSE2 |
BL21(DE3) | pGEX6P-2 | CuMTS3 |
BL21(DE3) | pGEX6P-2 | CuMTS62 |
Restriction Digest
Host | Sample | Enzyme | Buffer | |
---|---|---|---|---|
Vector | Insert | |||
BL21(DE3)PlysS | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
BL21(DE3)PlysS | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
BL21(DE3)PlysS | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
BL21(DE3)PlysS | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
BL21(DE3)PlysS | pGEX6P-2 | CuMTSE1 | Sal1, Not1 | Buffer H |
BL21(DE3)PlysS | pGEX6P-2 | CuMTSE2 | Sal1, Not1 | Buffer H |
BL21(DE3)PlysS | pGEX6P-2 | CuMTS3 | Sal1, Not1 | Buffer H |
BL21(DE3)PlysS | pGEX6P-2 | CuMTS62 | Sal1, Not1 | Buffer H |
June 11, 2014
KOBAYASHI
AGE
Lane | Sample/Marker | Enzyme | |
---|---|---|---|
Vector | Insert | ||
1 | λDNA-HindⅢ | ||
2 | pGEX6P-2 | CuMTSE1 | N.D |
3 | pGEX6P-2 | CuMTSE1 | EcoR1 |
4 | pGEX6P-2 | CuMTSE1 | Sal1, Not1 |
5 | pGEX6P-2 | CuMTSE2 | N.D |
6 | pGEX6P-2 | CuMTSE2 | EcoR1 |
7 | pGEX6P-2 | CuMTSE2 | Sal1, Not1 |
8 | pGEX6P-2 | CuMTS3 | N.D |
9 | pGEX6P-2 | CuMTS3 | EcoR1 |
10 | pGEX6P-2 | CuMTS62 | N.D |
11 | pGEX6P-2 | CuMTS62 | EcoR1 |
12 | pGEX6P-2 | CuMTS62 | Sal1, Not1 |
June 12, 2014
KOBAYASHI
Transformation
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 |
June 13, 2014
KOBAYASHI
Colony Isolation
Select 6 colonies per plate from 4 plates from June 12.
Host | Sample | Section | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | 1-a, 1-b |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | 2-a, 2-b |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 | 3-a, 3-b |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 | 6-a, 6-b |
June 14, 2014
ONISHI
Miniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
1-a | pGEX6P-2 | CuMTSE1 |
1-b | pGEX6P-2 | CuMTSE1 |
2-a | pGEX6P-2 | CuMTSE2 |
2-b | pGEX6P-2 | CuMTSE2 |
3-a | pGEX6P-2 | CuMTS3 |
3-b | pGEX6P-2 | CuMTS3 |
6-a | pGEX6P-2 | CuMTS62 |
6-b | pGEX6P-2 | CuMTS62 |
Miniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3) | pGEX6P-2 | CuMTSE1 |
BL21(DE3) | pGEX6P-2 | CuMTSE2 |
BL21(DE3) | pGEX6P-2 | CuMTS3 |
BL21(DE3) | pGEX6P-2 | CuMTS62 |
Restriction Digest
Host | Sample | Enzyme | Buffer | |
---|---|---|---|---|
Vector | Insert | |||
1-a | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
1-b | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
2-a | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
2-b | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
3-a | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
3-b | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
6-a | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
6-b | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
AGE
Lane | Sample | Insert | Enzyme | Buffer | ||
---|---|---|---|---|---|---|
Marker | Host | Vector | ||||
1 | 1kb Ladder | |||||
2 | λDNA-HindⅢ | |||||
3 | 1-a | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H | |
4 | 1-b | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H | |
5 | 2-a | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H | |
6 | 2-b | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H | |
7 | 3-a | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H | |
8 | 3-b | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H | 9 | 6-a | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
10 | 6-b | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H | |
11 | BL21(DE3) | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H | |
12 | BL21(DE3) | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H | |
13 | BL21(DE3) | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H | |
14 | BL21(DE3) | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
Applied λDNA-HindⅢ to compare and contrast AGE of July 14 and June 11.
June 16, 2014
YAMAJI
Preculture
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 |
BL21(DE3)pLysS | pGEX6P-2 |
Main Culture
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced |
June 18, 2014
YAMAJI
Preculture
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
Main Culture
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
Protein Extraction (E.coli)
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
Preparations for SDS-PAGE
Add SDS sample buffer to protein extractions in a flask, incubate at 37℃ for 15 minutes and keep it at 4℃.
June 19, 2014
KITOMI
SDS-PAGE
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
June 21, 2014
SOGA
Miniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
PCR
Vector | Sample | DNA Polymerase | |
---|---|---|---|
Insert | Primer | ||
pGEX6P-2 | CuMTSE1 | P-F1, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE2 | P-F2, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE1 | P-F1, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE2 | P-F2, P-R | KOD-FX-NEO |
AGE
Lane | Sample | Insert | Primer | ||
---|---|---|---|---|---|
Marker | Host | Vector | |||
1 | 1kb Ladder | ||||
2 | BY4247 | p427-TEF | |||
3 | pGEX6P-2 | CuMTSE1 | P-F1, P-R | ||
4 | pGEX6P-2 | CuMTSE2 | P-F2, P-R | ||
5 | pGEX6P-2 | CuMTSE1 | P-F1, P-R | ||
6 | pGEX6P-2 | CuMTSE2 | P-F2, P-R |
June 24, 2014
SOGA
Preculture
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | ||
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
June 25, 2014
KOBAYASHI
Cultivated following samples in Big Scale at 30℃ overnight in shaken culture.
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | ||
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
DNA Refinement1
Sample | |
---|---|
DNA | Primer |
CuMTSE1 | P-F1, P-R |
CuMTSE2 | P-F2, P-R |
Restriction Digest
Vector | Sample | Enzyme | Buffer | |
---|---|---|---|---|
DNA | Primer | |||
CuMTSE1 | P-F1, P-R | Spe1, Xho1 | Buffer H | |
CuMTSE2 | P-F2, P-R | Spe1, Xho1 | Buffer H | |
p427-TEF | Spe1, Xho1 | Buffer H |
DNA Refinement2
Vector | Sample | Enzyme | |
---|---|---|---|
DNA | Primer | ||
CuMTSE1 | P-F1, P-R | Spe1, Xho1 | |
CuMTSE2 | P-F2, P-R | Spe1, Xho1 | |
p427-TEF | Spe1, Xho1 |
Main Culture
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
Ligation
Sample | ||
---|---|---|
Vector | DNA | Primer |
p427-TEF | CuMTSE1 | P-F1, P-R |
p427-TEF | CuMTSE1 | P-F2, P-R |
p427-TEF | CuMTSE2 | P-F1, P-R |
p427-TEF | CuMTSE2 | P-F2, P-R |
Transformation
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
DH5α | p427-TEF | CuMTSE1 | P-F1, P-R |
DH5α | p427-TEF | CuMTSE1 | P-F2, P-R |
DH5α | p427-TEF | CuMTSE2 | P-F1, P-R |
DH5α | p427-TEF | CuMTSE2 | P-F2, P-R |
June 26, 2014
SAWANO
Colony Isolation
Isolate each colony from the plates which were inoculated on June 25.
Host | Sample | Section | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | p427-TEF | CuMTSE1 | P-F1, P-R | 1~4 |
DH5α | p427-TEF | CuMTSE1 | P-F2, P-R | a~h |
DH5α | p427-TEF | CuMTSE1 | P-F2, P-R | ア~タ |
June 27, 2014
SHIMADA
Colony Sweep
Lane | Sample | ||
---|---|---|---|
Host | Vector | Insert | |
1 | BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
2 | 1 | ||
3 | 2 | ||
4 | 3 | ||
5 | 4 | ||
6 | A | ||
7 | B | ||
8 | C | ||
9 | D | ||
10 | E | ||
11 | F | ||
12 | G | ||
13 | H |
Colony Sweep
Lane | Sample | ||
---|---|---|---|
Host | Vector | Insert | |
1 | BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
2 | ア | ||
3 | イ | ||
4 | ウ | ||
5 | エ | ||
6 | オ | ||
7 | カ | ||
8 | キ | ||
9 | ク | ||
10 | ケ | ||
11 | コ | ||
12 | サ | ||
13 | シ | ||
14 | ス | ||
15 | セ | ||
16 | ソ | ||
17 | タ |
Identified targeted genes in 4, c, h, イ, カ, セ
Miniprep
Sample | |
---|---|
Host | |
4 | |
C | |
H | |
イ | |
カ | |
セ |
Restriction Digest
Sample | Enzyme | Buffer |
---|---|---|
Host | ||
4 | Spe1, Xho1 | Buffer H |
C | Spe1, Xho1 | Buffer H |
H | Spe1, Xho1 | Buffer H |
イ | Spe1, Xho1 | Buffer H |
カ | Spe1, Xho1 | Buffer H |
AGE
Lane | Sample | Primer | Enzyme | ||
---|---|---|---|---|---|
Marker | Host | Insert | |||
1 | 1kb Ladder | ||||
2 | CuMTSE1 | P-F1, P-R | |||
3 | 4 | Spe1, Xho1 | |||
4 | 4 | Spe1, Xho1 | |||
5 | 4 | N.D | |||
6 | CuMTSE1 | P-F2, P-R | |||
7 | c | Spe1, Xho1 | |||
8 | c | Spe1, Xho1 | |||
9 | c | N.D | |||
10 | h | Spe1, Xho1 | |||
11 | h | Spe1, Xho1 | |||
12 | h | N.D |
Transformation (electroportation)
Host | Sample | |||
---|---|---|---|---|
Vector | Insert | Primer | ||
BY4247 | p427-TEF | CuMTSE1 | P-F1, P-R | |
BY4247 | p427-TEF | CuMTSE1 | P-F2, P-R |
June 30, 2014
SOGA
Inoculation
Divide each of 2 plates into 8 sections and inoculate following samples on them.
Host | Sample | Section | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
BY4247 | p427-TEF | CuMTSE1 | P-F1, P-R | A~H |
BY4247 | p427-TEF | CuMTSE1 | P-F2, P-R | 一~ハ |
July
July 1, 2014
KOBAYASHI
Protein Extraction (Saccharomyces cerevisiae)
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | CuMTSE1 | P-F1, P-R |
BY4247 | p427-TEF | CuMTSE1 | P-F2, P-R |
July 2, 2014
ONISHI
Miniprep
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTSE1 | P-F1, P-R |
BY4247 | p427-TEF | CuMTSE1 | P-F2, P-R |
Restriction Digest
Sample | Enzyme | Buffer | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
p427-TEF | CuMTSE1 | EcoR1 | Buffer H | |
p427-TEF | CuMTSE1 | P-F1, P-R | EcoR1 | Buffer H |
p427-TEF | CuMTSE1 | P-F2, P-R | EcoR1 | Buffer H |
p427-TEF | CuMTSE1 | Sac1, Kpn1 | Buffer L | |
p427-TEF | CuMTSE1 | P-F1, P-R | Sac1, Kpn1 | Buffer L |
p427-TEF | CuMTSE1 | P-F2,P-R | Sac1, Kpn1 | Buffer L |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Vector | Insert | Primer | ||
1 | p427-TEF | N.D | |||
2 | p427-TEF | CuMTSE1 | P-F1, P-R | N.D | |
3 | p427-TEF | CuMTSE1 | P-F2, P-R | N.D | |
4 | 1kb Ladder | ||||
5 | p427-TEF | EcoR1 | |||
6 | p427-TEF | CuMTSE1 | P-F1, P-R | EcoR1 | |
7 | p427-TEF | CuMTSE1 | P-F2, P-R | EcoR1 | |
8 | 1kb Ladder | ||||
9 | p427-TEF | Kpn1 | |||
10 | p427-TEF | CuMTSE1 | P-F1, P-R | Kpn1 | |
11 | p427-TEF | CuMTSE1 | P-F2, P-R | Kpn1 | |
12 | 1kb Ladder | ||||
13 | p427-TEF | Sac1 | |||
14 | p427-TEF | CuMTSE1 | P-F1, P-R | Sac1 | |
15 | p427-TEF | CuMTSE1 | P-F2, P-R | Sac1 | |
Restriction Digest
Sample | Enzyme | Buffer | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
p427-TEF | CuMTSE1 | EcoR1 | Buffer H | |
p427-TEF | CuMTSE1 | P-F1, P-R | EcoR1 | Buffer H |
p427-TEF | CuMTSE1 | P-F2, P-R | EcoR1 | Buffer H |
p427-TEF | CuMTSE1 | Sac1 | Buffer L | |
p427-TEF | CuMTSE1 | P-F1, P-R | Sac1 | Buffer L |
p427-TEF | CuMTSE1 | P-F2,P-R | Sac1 | Buffer L |
p427-TEF | CuMTSE1 | Kpn1 | Buffer L | |
p427-TEF | CuMTSE1 | P-F1, P-R | Kpn1 | Buffer L |
p427-TEF | CuMTSE1 | P-F2, P-R | Kpn1 | Buffer L |
p427-TEF | CuMTSE1 | Spe1, Xho1 | Buffer H | |
p427-TEF | CuMTSE1 | P-F1, P-R | Spe1, Xho1 | Buffer H |
p427-TEF | CuMTSE1 | P-F2, P-R | Spe1, Xho1 | Buffer H |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Vector | Insert | Primer | ||
1 | 1kb Ladder | ||||
2 | p427-TEF | CuMTSE1 | EcoR1 | ||
3 | p427-TEF | CuMTSE1 | P-F1, P-R | EcoR1 | |
4 | p427-TEF | CuMTSE1 | P-F2, P-R | EcoR1 | |
5 | p427-TEF | CuMTSE1 | Kpn1 | ||
6 | p427-TEF | CuMTSE1 | P-F1, P-R | Kpn1 | |
7 | p427-TEF | CuMTSE1 | P-F2, P-R | Kpn1 | |
8 | p427-TEF | CuMTSE1 | Sac1 | ||
9 | p427-TEF | CuMTSE1 | P-F1, P-R | Sac1 | |
10 | p427-TEF | CuMTSE1 | P-F2, P-R | Sac1 | |
11 | p427-TEF | CuMTSE1 | Spe1, Xho1 | ||
12 | p427-TEF | CuMTSE1 | P-F1, P-R | Spe1, Xho1 | |
13 | p427-TEF | CuMTSE1 | P-F2, P-R | Spe1, Xho1 |
July 3, 2014
OKAMOTO
PCR
Sample | DNA Polymerase | ||
---|---|---|---|
Vector | Insert | Primer | |
pGEX6P-2 | CuMTSE2 | P-F1, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE2 | P-F2, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTS3 | P-F1, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTS3 | P-F2, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTS62 | P-F1, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTS62 | P-F2, P-R | KOD-FX-NEO |
Protein Extraction (Saccharomyces cerevisiae)
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
六 | p427-TEF | CuMTSE1 | P-F1, P-R |
A | p427-TEF | CuMTSE1 | P-F2, P-R |
July 4, 2014
SHIMADA
AGE
Lane | Sample/Marker | |||
---|---|---|---|---|
Marker | Vector | Insert | Primer | |
1 | 1kb Ladder | |||
2 | pGEX6P-2 | CuMTSE2 | P-F1, P-R | |
3 | pGEX6P-2 | 1kb Ladder+K83:N91 | P-F2, P-R | |
4 | pGEX6P-2 | CuMTS3 | P-F1, P-R | |
5 | pGEX6P-2 | CuMTS3 | P-F2, P-R | |
6 | pGEX6P-2 | CuMTS62 | P-F1, P-R | |
7 | pGEX6P-2 | CuMTS62 | P-F2, P-R |
PCR
Sample | DNA Polymerase | ||
---|---|---|---|
Vector | Insert | Primer | |
pGEX6P-2 | CuMTSE2 | P-F1, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE2 | P-F2, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTS3 | P-F1, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTS3 | P-F2, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTS62 | P-F1, P-R | KOD-FX-NEO |
pGEX6P-2 | CuMTS62 | P-F2, P-R | KOD-FX-NEO |
AGE
Lane | Sample | |||
---|---|---|---|---|
Marker | Vector | Insert | Primer | |
1 | 1kb Ladder | |||
2 | pGEX6P-2 | CuMTSE2 | P-F1, P-R | |
3 | pGEX6P-2 | 1kb Ladder+K83:N91 | P-F2, P-R | |
4 | pGEX6P-2 | CuMTS3 | P-F1, P-R | |
5 | pGEX6P-2 | CuMTS3 | P-F2, P-R | |
6 | pGEX6P-2 | CuMTS62 | P-F1, P-R | |
7 | pGEX6P-2 | CuMTS62 | P-F2, P-R |
July 5, 2014
KOBAYASHI
SDS-PAGE
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BL21(DE3)pLysS | pGEX6P-2 | ||
4 | pGEX6P-2 | CuMTSE1 | P-F1, P-R |
C | pGEX6P-2 | CuMTSE2 | P-F2, P-R |
BY4247 | p427-TEF | ||
六 | p427-TEF | CuMTSE1 | P-F1, P-R |
A | p427-TEF | CuMTSE2 | P-F2, P-R |
July 6, 2014
SOGA
DNA Refinement1
Sample | |
---|---|
DNA | Primer |
CuMTSE1 | P-F1, P-R |
CuMTSE2 | P-F2, P-R |
CuMTS3 | P-F1, P-R |
CuMTS62 | P-F1, P-R |
Restriction Digest
Vector | Sample | Enzyme | Buffer | |
---|---|---|---|---|
DNA | Primer | |||
CuMTSE2 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer | |
CuMTSE2 | P-F2, P-R | Spe1, Xho1 | Fast digest buffer | |
CuMTS3 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer | CuMTS62 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer | p427-TEF | Spe1, Xho1 | Fast digest buffer |
DNA Refinement2
Sample | Enzyme | |
---|---|---|
DNA | Primer | |
CuMTSE2 | P-F1, P-R | Spe1, Xho1 |
CuMTSE2 | P-F2, P-R | Spe1, Xho1 |
Ligation
Vector | Sample | Enzyme | |
---|---|---|---|
DNA | Primer | ||
p427-TEF | CuMTSE2 | P-F1, P-R | Spe1, Xho1 |
p427-TEF | CuMTSE2 | P-F2, P-R | Spe1, Xho1 |
DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.)
Transformation (E.coli)
Host | Sample | Enzyme | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | p427-TEF | CuMTSE2 | P-F1, P-R | Spe1, Xho1 |
DH5α | p427-TEF | CuMTSE2 | P-F2, P-R | Spe1, Xho1 |
DH5α | p427-TEF | CuMTS3 | P-F1, P-R | Spe1, Xho1 |
DH5α | p427-TEF | CuMTS62 | P-F1, P-R | Spe1, Xho1 |
July 7, 2014
KOBAYASHI
Colony Isolation
Isolate each colony from the plates which were inoculated on July 6.
Host | Sample | Section | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | p427-TEF | CuMTSE2 | P-F1, P-R | 1~16 |
DH5α | p427-TEF | CuMTS3 | P-F1, P-R | ア~エ |
DH5α | p427-TEF | CuMTS62 | P-F1, P-R | a~p |
Transformation (E.coli)
Host | Sample | Enzyme | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | p427-TEF | CuMTSE2 | P-F2, P-R | Spe1, Xho1 |
July 8, 2014
OKAMOTO
Colony Sweep
Lane | Sample | |
---|---|---|
Host | ||
1 | DH5α | p427-TEF |
2 | a | |
3 | b | |
4 | c | |
5 | d | |
6 | e | |
7 | f | |
8 | g | |
9 | h | |
10 | i | |
11 | j | |
12 | k | |
13 | l | |
14 | m | |
15 | n | |
16 | o | |
17 | p |
Colony Sweep
Lane | Sample | |
---|---|---|
Host | ||
1 | DH5α | p427-TEF |
2 | ア | |
3 | イ | |
4 | ウ | |
5 | エ |
Colony Sweep
Lane | Sample | |
---|---|---|
Host | ||
1 | DH5α | p427-TEF |
2 | 1 | |
3 | 2 | |
4 | 3 | |
5 | 4 | |
6 | 5 | |
7 | 6 | |
8 | 7 | |
9 | 8 | |
10 | 9 | |
11 | 10 | |
12 | 11 | |
13 | 12 | |
14 | 13 | |
15 | 14 | |
16 | 15 | |
17 | 16 |
July 9, 2014
OKAMOTO
Main Culture
Sample |
---|
Host |
d |
h |
l |
ア |
イ |
エ |
Mini Prep
Sample |
---|
Host |
D |
H |
L |
ア |
イ |
エ |
Restriction Digest
Vector | Sample | Enzyme | Buffer | |
---|---|---|---|---|
Insert | Primer | |||
p427-TEF(d) | CuMTS3 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(h) | CuMTS3 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(l) | CuMTS3 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(ア) | CuMTS62 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(イ) | CuMTS62 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(エ) | CuMTS62 | P-F1, P-R | Spe1, Xho1 | Fast digest buffer |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Vector | Insert | Primer | ||
1 | 1kb Ladder | ||||
2 | p427-TEF(ア) | CuMTS3 | P-F1, P-R | N.D | |
3 | p427-TEF(ア) | CuMTS3 | P-F1, P-R | Spe1, Xho1 | |
4 | p427-TEF(イ) | CuMTS3 | P-F1, P-R | N.D | |
5 | p427-TEF(イ) | CuMTS3 | P-F1, P-R | Spe1, Xho1 | |
6 | p427-TEF(エ) | CuMTS3 | P-F1, P-R | N.D | |
7 | p427-TEF(エ) | CuMTS3 | P-F1, P-R | Spe1, Xho1 | |
8 | p427-TEF(d) | CuMTS62 | P-F1, P-R | N.D | |
9 | p427-TEF(d) | CuMTS62 | P-F1, P-R | Spe1, Xho1 | |
10 | p427-TEF(h) | CuMTS62 | P-F1, P-R | N.D | |
11 | p427-TEF(h) | CuMTS62 | P-F1, P-R | Spe1, Xho1 | |
12 | p427-TEF(l) | CuMTS62 | P-F1, P-R | N.D | |
13 | p427-TEF(l) | CuMTS62 | P-F1, P-R | Spe1, Xho1 |
August
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