CII
7.24
PCR
Yufeng Mo
Name | Total |
CII 1 | 50 μl |
CII 2 | 50 μl |
CII control group | 30 μl |
7.25
Restriction Enzyme Digestion
Yufeng Mo
Name | Enzyme |
CII | XbaI and PstI |
7.26
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
CII | psb1c3(Xba I and Pst I cut) | DH5α | LB(chl added) |
7.27(29,30?)
Colony PCR
Yufeng Mo
Number | Name |
1 | CII |
![](https://static.igem.org/mediawiki/2016/f/f8/T--HUST-China--notebook-CII-fig01.png)
CIII
7.14
PCR
7.15
Restriction Enzyme Digestion
Yufeng Mo
Name | Enzyme |
plasmid psb1c3 1 | XbaI and SpeI |
plasmid psb1c3 2 | XbaI and SpeI |
![](https://static.igem.org/mediawiki/2016/8/8c/T--HUST-China--notebook-CIII-fig01.png)
7.17
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
CIII | psb1c3(XbaI and SpeI cut) | DH5α | LB(chl added) |
7.18
PCR
Yufeng Mo
Number | Name |
1 | CIII 1 |
2 | CIII 2 |
3 | CIII 3 |
4 | CIII 4 |
5 | CIII 5 |
6 | CIII 6 |
![](https://static.igem.org/mediawiki/2016/c/ca/T--HUST-China--notebook-CIII-fig02.png)
RBS-CII
PCR
more details,protocol
Yufeng Mo
Name | Total |
RBS-CII 2 | 40 μl |
RBS-CII 1 | 40 μl |
RBS-CII 0 | 20 μl |
9.4
Restriction Enzyme Digestion
Yufeng Mo
Name | Enzyme |
RBS-CII | XbaI and PstI |
9.4
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
RBS-CII | psb1c3(Xba I and Pst I cut) | DH5α | LB(chl added) |
9.5
Colony PCR
Yufeng Mo
Number | Name |
1 | RBS-CII 1 |
2 | RBS-CII 2 |
![](https://static.igem.org/mediawiki/2016/c/ca/T--HUST-China--notebook-RBS-CII-fig01.png)
RBS-CIII
8.7
PCR
Ziyi Jiang
Name | Total |
RBS-CIII | 50 μl |
![](https://static.igem.org/mediawiki/2016/5/58/T--HUST-China--notebook-RBS-CIII-fig01.png)
8.7
Restriction Enzyme Digestion
Ziyi Jiang
Name | Enzyme |
psb1c3 | XbaI and PstI |
RBS-CIII | XbaI and PstI |
8.10
DNA ligation&transformation
Zhenyi Guo
Number | Name |
2 | CIII-RBS |
![](https://static.igem.org/mediawiki/2016/9/94/T--HUST-China--notebook-RBS-CIII-fig02.png)
RBS-CIII-RBS-CIII
8.18
Restriction Enzyme Digestion
Ziyi Jiang
Name | Enzyme |
RBS-CIII | EcoRI and SpeI |
RBS-CIII | EcoRI and XbaI |
![](https://static.igem.org/mediawiki/2016/6/67/T--HUST-China--notebook-RBS-CIII-RBS-CIII-fig01.png)
8.18
DNA ligation&Transformation
Ziyi Jiang
Name | Backbone | Strain | Medium |
CIII-RBS | CIII-RBS-psb1c3(EcoRI and XbaI cut) | DH5α | LB(chl added) |
8.19
Colony PCR
Successfully align the sequencing results with the objective sequences:
Ziyi Jiang
Number | Name |
13 | RBS-CIII-RBS-CIII-psb1c3 |
19 | RBS-CIII-RBS-CIII-psb1c3 |
![](https://static.igem.org/mediawiki/2016/6/67/T--HUST-China--notebook-RBS-CIII-RBS-CIII-fig02.png)
RBS-CII-RBS-CII
9.12
PCR
Yufeng Mo
Name | Total |
RBS-CII | 50 μl |
9.14
Restriction Enzyme Digestion
Yufeng Mo
Name | Enzyme |
RBS-CII | EcoRI and XbaI |
RBS-CII-psb1c3 | EcoRI and SpeI |
9.14
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
RBS-CII | RBS-CII-psb1c3(EcoRI and SpeI cut) | DH5α | LB(chl added) |
9.15
Colony PCR
Successfully align the sequencing results with the objective sequences:
Zhi Zeng
Number | Name |
1 | RBS-CII-RBS-CII-psb1c3 |
![](https://static.igem.org/mediawiki/2016/e/e2/T--HUST-China--notebook-RBS-CII-RBS-CII-fig01.png)
pRM-RBS-GFP-LAVtag
7.18
PCR
Xiaoyang Li
Name | Total |
GFP-F 1 | 50 μl |
Name | Total |
GFP-X-F | 30 μl |
GFP-X-F | 30 μl |
GFP-X-F | 30 μl |
![](https://static.igem.org/mediawiki/2016/b/b7/T--HUST-China--notebook-pRM-RBS-GFP-LVAtag-fig01.png)
7.19
Restriction Enzyme Digestion
Name | Enzyme |
GFP | EcoRI and SpeI |
psb1c3 | EcoRI and SpeI |
7.20
DNA ligation&transformation
Xiaoyang Li
Name | Backbone | Strain | Medium |
pRM-RBS-GFP | psb1c3(EcoRI and SpeI cut) | DH5α | LB(chl added) |
7.24
Colony PCR
Xiaoyang Li
Number | Name |
1 | pRM-RBS-GFP-LVAtag |
![](https://static.igem.org/mediawiki/2016/e/ec/T--HUST-China--notebook-pRM-RBS-GFP-LVAtag-fig02.png)
8.21
Yufeng Mo
Name | Enzyme |
pRM-RBS-GFP-LVAtag | XbaI and PstI |
Duet-pet | XbaI and PstI |
8.21
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
pRM-RBS-GFP-LVAtag | pet-Duet-1(XbaI and PstI cut) | DH5α | LB(Amp added) |
8.22
Colony PCR
Yufeng Mo
Number | Name |
1 | pRM-RBS-GFP-LVAtag-Duet-pet 1 |
3 | pRM-RBS-GFP-LVAtag-Duet-pet 3 |
4 | pRM-RBS-GFP-LVAtag-Duet-pet 4 |
5 | pRM-RBS-GFP-LVAtag-Duet-pet 5 |
6 | pRM-RBS-GFP-LVAtag-Duet-pet 6 |
![](https://static.igem.org/mediawiki/2016/6/66/T--HUST-China--notebook-pRM-RBS-GFP-LVAtag-fig03.png)
7.24
Colony PCR
Xiaoyang Li
Number | Name |
1 | pRM-RBS-GFP-LVAtag |
![](https://static.igem.org/mediawiki/2016/e/ec/T--HUST-China--notebook-pRM-RBS-GFP-LVAtag-fig02.png)
pRE-RBS-GFP-LVAtag
8.23
Restriction Enzyme Digestion
Yufeng Mo
Name | Enzyme |
pRE-RBS-GFP-LVAtag | XbaI and PstI |
8.23
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
pRE-RBS-GFP-LVAtag | psb1c3(XbaI and PstI cut) | DH5α | LB(chl added) |
8.24
Colony PCR
Yufeng Mo
Number | Name |
1 | pRE-RBS-GFP-LVAtag-psb1c3 |
![](https://static.igem.org/mediawiki/2016/3/38/T--HUST-China--notebook-pRE-RBS-GFP-LVAtag-fig01.png)
8.29
Restriction Enzyme Digestion
Yufeng Mo,Zhenyi Guo
Name | Enzyme |
pRE-RBS-GFP-LVAtag-psb1c3 | EcoRI and PstI |
8.29
DNA ligation&transformation
Xiao Deng,Zhenyi Guo
Name | Backbone | Strain | Medium |
pRE-RBS-GFP-LVAtag | petDuet-1(EcoRI and PstI cut) | DH5α | LB(Amp added) |
8.30
Colony PCR
Yufeng Mo
Number | Name |
1 | pRE-RBS-GFP-LVAtag-petDuet-1 1 |
3 | pRE-RBS-GFP-LVAtag-petDuet-1 3 |
5 | pRE-RBS-GFP-LVAtag-petDuet-1 5 |
8 | pRE-RBS-GFP-LVAtag-petDuet-1 8 |
![](https://static.igem.org/mediawiki/2016/d/da/T--HUST-China--notebook-pRE-RBS-GFP-LVAtag-fig02.png)
pR-RBS-GFP-LAVtag
8.2
PCR
Zhenyi Guo
Name | Total |
pR-RBS-GFP-tag | 50 μl |
![](https://static.igem.org/mediawiki/2016/4/45/T--HUST-China--notebook-pR-RBS-GFP-LVAtag-fig01.png)
Restriction Enzyme Digestion
Zhenyi Guo
Name | Enzyme |
psb1c3 | EcoRI and PstI |
pR-RBS-GFP-tag | EcoRI and PstI |
![](https://static.igem.org/mediawiki/2016/b/b7/T--HUST-China--notebook-pR-RBS-GFP-LVAtag-fig02.png)
DNA ligation&Transformation
Zhenyi Guo
Name | Backbone | Strain | Medium |
pR-RBS-GFP-tag | psb1c3(EcoRI and PstI cut) | DH5α | LB(chl added) |
8.4
Colony PCR
Successfully align the sequencing results with the objective sequences:
Number | Name |
2 | pR-RBS-GFP-tag |
![](https://static.igem.org/mediawiki/2016/3/3d/T--HUST-China--notebook-pR-RBS-GFP-LVAtag-fig03.png)
8.24
Restriction Enzyme Digestion
Zhenyi Guo
Name | Enzyme |
pR-RBS-GFP-tag | EcoRI and PstI |
8.24
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
pR-RBS-GFP-tag | petDuet-1(XbaI and PstI cut) | DH5α | LB(Amp added) |
8.26
Colony PCR
Yufeng Mo
Number | Name |
4 | pR-RBS-GFP-tag-petDuet-1 4 |
6 | pR-RBS-GFP-tag-petDuet-1 6 |
7 | pR-RBS-GFP-tag-petDuet-1 7 |
![](https://static.igem.org/mediawiki/2016/9/9a/T--HUST-China--notebook-pR-RBS-GFP-LVAtag-fig04.png)
SnRK2.2
8.25
Site-mutated PCR
Fengyi Li
Name | Total |
SnRK2.2 | 40 μl |
![](https://static.igem.org/mediawiki/2016/4/4b/T--HUST-China--notebook-SnRK2.2-fig01.png)
9.10
PCR
more details, protocol
Yufeng Mo
Name | Total |
SnRK2.2 1 | 40 μl |
SnRK2.2 2 | 40 μl |
SnRK2.2 0 | 20 μl |
9.10
Restriction Enzyme Digestion
Yufeng Mo
Name | Enzyme |
SnRK2.2 | XbaI and PstI |
9.10
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
SnRK2.2 | psb1c3(XbaI and PstI cut) | DH5α | LB(chl added) |
9.12
Colony PCR
Successfully align the sequencing results with the objective sequences:
Yufeng Mo
Number | Name |
1 | SnRK2.2-psb1c3 |
![](https://static.igem.org/mediawiki/2016/5/58/T--HUST-China--notebook-SnRK2.2-fig02.png)
ABF2
9.10
PCR
more details,protocol
Yufeng Mo
Name | Total |
ABF2 1 | 40 μl |
ABF2 2 | 40 μl |
ABF2 0 | 20 μl |
9.10
Restriction Enzyme Digestion
Yufeng Mo
Name | Enzyme |
ABF2 | XbaI and PstI |
9.10
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
ABF2 | psb1c3(XbaI and PstI cut) | DH5α | LB(chl added) |
9.11
Colony PCR
Yufeng Mo
Number | Name |
2 | ABF2 2 |
3 | ABF2 3 |
4 | ABF2 4 |
![](https://static.igem.org/mediawiki/2016/8/80/T--HUST-China--notebook-ABF2-fig01.png)
PP2CA
7.25
PCR
Yufeng Mo
Name | Total |
PP2CA 1 | 25 μl |
PP2CA 2 | 25 μl |
PP2CA control group | 25 μl |
7.26
Restriction enzyme digestion
Yufeng Mo
Name | Enzyme |
PP2CA | XbaI and PstI |
7.26
DNA ligation&transformation
Yufeng Mo
Name | Backbone | Strain | Medium |
PP2CA | psb1c3(Xba I and Pst I cut) | E.coli DH5α | LB(chl added) |
7.27
Colony PCR
Yufeng Mo
Number | Name |
2 | PP2CA |
![](https://static.igem.org/mediawiki/2016/c/cd/T--HUST-China--notebook-PP2CA-fig01.png)
prd29A
9.27
PCR
Zhi Zeng
Name | Total |
prd29a | 50 μl |
![](https://static.igem.org/mediawiki/2016/7/7d/T--HUST-China--notebook-prd29a-fig01.png)
9.27
Restriction Enzyme Digestion
Zhenyi Guo
Name | Enzyme |
prd29a | XbaI and PstI |
![](https://static.igem.org/mediawiki/2016/0/05/T--HUST-China--notebook-prd29a-fig02.png)
9.28
Colony PCR
Successfully align the sequencing results with the objective sequences:
Zhenyi Guo
Name | Enzyme |
prd29a | XbaI and PstI |
![](https://static.igem.org/mediawiki/2016/0/05/T--HUST-China--notebook-prd29a-fig02.png)
9.27
Restriction Enzyme Digestion
Zhenyi Guo
Number | Name |
1 | prd29a-psb1c3 |
2 | prd29a-psb1c3 |
![](https://static.igem.org/mediawiki/2016/4/4a/T--HUST-China--notebook-prd29a-fig03.png)
CI-TT-pR-RBS-GFP-LVAtag&CII-TT-pRE-RBS-GFP-LVAtag
8.17
Restriction Enzyme Digestion
Lin Di
Name | Enzyme |
Duet-pet | EcoRI and PstI |
CI-TT-pR-RBS-GFP-LVAtag | EcoRI and PstI |
CII-TT-pRE-RBS-GFP-LVAtag | EcoRI and PstI |
![](https://static.igem.org/mediawiki/2016/d/df/T--HUST-China--notebook-CI-TT%26CII-TT-fig01.png)
![](https://static.igem.org/mediawiki/2016/b/be/T--HUST-China--notebook-CI-TT%26CII-TT-fig02.png)
8.17
DNA ligation&Transformation
Lin Di
Name | Backbone | Strain | Medium |
CI-TTADH1-pR-RBS-GFP-LVAtag | Duet-pet(T7) | DH5α | LB(Amp added) |
CII-TTADH1-pRE-RBS-GFP-LVAtag | Duet-pet(T7) | DH5α | LB(Amp added) |
8.18
Colony PCR
Successfully align the sequencing results with the objective sequences:
Lin Di
Number | Name |
6 | CI-TTADH1-pR-RBS-GFP-LVAtag-petDuet-1 6 |
5 | CII-TTADH1-pRE-RBS-GFP-LVAtag-petDuet-1 5 |
11 | CII-TTADH1-pRE-RBS-GFP-LVAtag -petDuet-1 11 |
![](https://static.igem.org/mediawiki/2016/9/91/T--HUST-China--notebook-CI-TT%26CII-TT-fig03.png)
![](https://static.igem.org/mediawiki/2016/0/02/T--HUST-China--notebook-CI-TT%26CII-TT-fig04.png)
Cro-TT-pRM-RBS-GFP-LVAtag-PSB1C3
8.14
PCR
Zhi Zeng
Name | Total |
Cro | 80 μl |
![](https://static.igem.org/mediawiki/2016/8/87/T--HUST-China--notebook-Cro-T-P-R-G-tag-fig01.png)
8.15
Restriction Enzyme Digestion
Zhi Zeng
Name | Enzyme |
pRM-RBS-GFP-LVAtag-psb1c3 | EcoRI and XbaI |
psb1c3 | XbaI and PstI |
![](https://static.igem.org/mediawiki/2016/b/b5/T--HUST-China--notebook-Cro-T-P-R-G-tag-fig02.png)
8.16
In-Fusion
Zhi Zeng
Insert | Vector | Total |
Cro&TT | pRM-RBS-GFP-LVAtag-PSB1C3(Cut by EcoRⅠ&XbaⅠ) | 5uL |
8.16
Transformation
Zhi Zeng
Name | Strain | Medium |
Cro-TT-pRM-RBS-GFP-LVAtag-PSB1C3 (ligate by Infusion) | DH5α | LB(Chl added) |
8.17
Colony PCR
Zhi Zeng
Name | Total |
Cro-TT-pRM-RBS-GFP-LVAtag-psb1c3 | 20uL |
![](https://static.igem.org/mediawiki/2016/3/3f/T--HUST-China--notebook-Cro-T-P-R-G-tag-fig03.png)
8.18
Restriction Enzyme Digestion
Zhi Zeng
Name | Enzyme |
Duet-pet | EcoRI and PstI |
Cro-TT-pRM-RBS-GFP-LVAtag | EcoRI and PstI |
8.18
DNA ligation&Transformation
Zhi Zeng
Name | Backbone | Strain | Medium |
Cro-TT-pRM-RBS-GFP-LVAtag | Duet-pet(T7) | DH5α | LB(Amp added) |
PP2CA-TTADH1
8.14
Restriction Enzyme Digestion
Ziyi Jiang
Name | Enzyme |
TTADH1-psb1c3 | EcoRI and XbaI |
PP2CA-psb1c3 | EcoRI and SpeI |
![](https://static.igem.org/mediawiki/2016/2/2b/T--HUST-China--notebook-PP-TT-fig01.png)
8.16
Colony PCR
Successfully align the sequencing results with the objective sequences:
Ziyi Jiang
Number | Name |
5 | PP2CA-TTADH1-psb1c3 5 |
7 | PP2CA-TTADH1-psb1c3 7 |
12 | PP2CA-TTADH1-psb1c3 12 |
![](https://static.igem.org/mediawiki/2016/4/4a/T--HUST-China--notebook-PP-TT-fig02.png)
PP2CA-TTADH1-pCyc
8.22
Overlap PCR
Zhi Zeng
Name | Total |
PP2CA-TTADH1-Pcyc | 80uL |
![](https://static.igem.org/mediawiki/2016/b/b2/T--HUST-China--notebook-PP-TT-pcyc-fig01.png)
8.22
Restriction Enzyme Digestion
Zhi Zeng
Backbone | Enzyme |
psb1c3 | PstI and XbaI |
![](https://static.igem.org/mediawiki/2016/5/54/T--HUST-China--notebook-PP-TT-pcyc-fig02.png)
8.23
In-Fusion
Zhi Zeng
Insert | Vector | Total |
PP2CA-TTADH1-pCyc | psb1c3(Cut by PstⅠ&XbaⅠ) | 5uL |
8.23
Transformation
Zhi Zeng
Name | Strain | Medium |
PP2CA-TTADH1-pCyc-psb1c3 (ligate by In-Fusion) | DH5α | LB(Chl added) |
8.24
Colony PCR
Zhi Zeng
Number | Name |
3 | PP2CA-TTADH1-pCyc-psb1c3 |
4 | PP2CA-TTADH1-pCyc-psb1c3 |
7 | PP2CA-TTADH1-pCyc-psb1c3 |
13 | PP2CA-TTADH1-pCyc-psb1c3 |
14 | PP2CA-TTADH1-pCyc-psb1c3 |
![](https://static.igem.org/mediawiki/2016/2/21/T--HUST-China--notebook-PP-TT-pcyc-fig03.png)
Kozak-SnRK2.2-TTADH1-pCyc
8.30
Overlap PCR
Name | Total |
Kozak-Snrk2.2-TTADH1-pCyc | 80uL |
![](https://static.igem.org/mediawiki/2016/4/41/T--HUST-China--notebook-K-S-T-P-fig02.png)
9.5
In-Fusion
Zhi Zeng
Insert | Vector | Total |
Kozak-Snrk2.2-TTADH1-pCyc | psb1c3(Cut by PstⅠ&XbaⅠ) | 5uL |
![](https://static.igem.org/mediawiki/2016/3/3b/T--HUST-China--notebook-K-S-T-P-fig01.png)
9.5
Transformation
Zhi Zeng
Name | Strain | Medium |
Kozak-Snrk2.2-TTADH1-pCyc-psb1c3(ligate by In-Fusion) | DH5α | LB(Chl added) |
9.7
Colony PCR
Zhi Zeng
Name | Total |
Kozak-Snrk2.2-TTADH1-pCyc-psb1c3 | 20uL |
![](https://static.igem.org/mediawiki/2016/5/50/T--HUST-China--notebook-K-S-T-P-fig03.png)
placm-pRE-RBS-Cro-RBS-CII-TT
9.6
Overlap PCR
Jiyue Qin
Name | Total |
Placm-pRE-RBS-Cro | 50uL |
9.7
In-Fusion
Jiyue Qin
Insert | Vector | Total |
Placm-pRE-RBS-Cro&RBS-cII-TT | psb1c3(Cut by PstⅠ&XbaⅠ) | 10uL |
9.6
Transformation
Jiyue Qin
Name | Strain | Medium |
Placm-pRE-RBS-Cro-RBS-CII-TT-psb1c3(ligate by In-Fusion) | DH5α | LB(Chl added) |
9.8
Colony PCR
Jiyue Qin
Number | Name |
1 | Placm-pRE-RBS-Cro-RBS-CII-TT-psb1c3 |
![](https://static.igem.org/mediawiki/2016/6/61/T--HUST-China--notebook-placm-p-R-Cro-fig01.png)
patp2-RBS-CI-TTADH1-pR-RBS-CIII
9.2
Overlap PCR
Jiyue Qin
Name | Total |
Patp2-RBS-CI-TT | 50uL |
![](https://static.igem.org/mediawiki/2016/e/e2/T--HUST-China--notebook-patp2-R-CI-TT-fig01.png)
9.3
In-Fusion
Jiyue Qin
Insert | Vector | Total |
Patp2-RBS-CI-TT&pR-RBS-CIII | psb1c3(Cut by PstⅠ&XbaⅠ) | 10uL |
![](https://static.igem.org/mediawiki/2016/e/e2/T--HUST-China--notebook-patp2-R-CI-TT-fig01.png)
9.3
Transformation
Jiyue Qin
Name | Strain | Medium |
Patp2-RBS-CI-TT-pR-RBS-CIII-psb1c3 | DH5α | LB(Chl added) |
9.4
Colony PCR
Jiyue Qin
Number | Name |
1 | Patp2-RBS-CI-TT-pR-RBS-CIII-psb1c3 |
![](https://static.igem.org/mediawiki/2016/4/4b/T--HUST-China--notebook-patp2-R-CI-TT-fig02.png)
RBS-iLDH-TT-pRM-RBS-beta-galactosidase
9.1
Restriction Enzyme Digestion
Jiyue Qin
Name | Enzyme |
GFP-LVAtag-psb1c3 | XbaI and PstI |
![](https://static.igem.org/mediawiki/2016/4/49/T--HUST-China--notebook-R-iLDH-T-p-R-β-fig01.png)
9.1
In-Fusion
Jiyue Qin
Insert | Vector | Total |
RBS-iLDH-TT&pRM-RBS-beta-galactosidase | psb1c3(Cut by PstⅠ&XbaⅠ) | 10uL |
9.1
Transformation
Jiyue Qin
Name | Strain | Medium |
RBS-iLDH-TT-pRM-RBS-beta-galactosidase-psb1c3 | DH5α | LB(Chl added) |
9.2
Colony PCR
Jiyue Qin
Number | Name |
1 | RBS-iLDH-TT-pRM-RBS-beta-galactosidase-psb1c3 |
![](https://static.igem.org/mediawiki/2016/d/d1/T--HUST-China--notebook-Whole_circuit_of_app-fig01.png)
plac-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-iLDH-TT-pRM-RBS-beta-galactosidase
9.7
In-Fusion
Zhenyi Guo
Insert | Vector | Total |
plac-pRE-RBS-Cro-RBS-CII-TT&patp2-RBS-CI-TT-pR-RBS-CIII-RBS-iLDH-TT-pRM-RBS-beta-galactosidase | psb1c3(Cut by PstⅠ&XbaⅠ) | 10uL |
9.7
Transformation
Zhenyi Guo
Name | Strain | Medium |
plac-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-iLDH-TT-pRM-RBS-beta-galactosidase | DH5α | LB(Chl added) |
9.9
Colony PCR
Zhenyi Guo
Number | Name |
1 | plac-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-iLDH-TT-pRM-RBS-beta-galactosidase |
![](https://static.igem.org/mediawiki/2016/d/d1/T--HUST-China--notebook-Whole_circuit_of_app-fig01.png)
9.11
Restriction Enzyme Digestion
Zhenyi Guo
Name | Enzyme |
Duet-pet | EcoRI and PstI |
plac-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-iLDH-TT-pRM-RBS-beta-galactosidase -psb1c3 | EcoRI and PstI |
9.7
Transformation
Zhenyi Guo
Name | Strain | Medium |
plac-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-iLDH-TT-pRM-RBS-beta-galactosidase | DH5α | LB(Chl added) |
9.17
DNA ligation&Transformation
Zhenyi Guo
Name | Backbone | Strain | Medium |
plac-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-iLDH-TT-pRM-RBS-beta-galactosidase | Duet-pet(T7)(cut by EcoRI and PstI) | DH5α | LB(Amp added) |
9.18
Colony PCR
Successfully align the sequencing results with the objective sequences:
Zhenyi Guo
Number | Name |
1 | plac-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-iLDH-TT-pRM-RBS-beta-galactosidase-Duet-pet |
![](https://static.igem.org/mediawiki/2016/c/c5/T--HUST-China--notebook-Whole_circuit_of_app-fig02.png)