Difference between revisions of "Team:HUST-China/Notebook"

Line 1,703: Line 1,703:
 
                         <img src="https://static.igem.org/mediawiki/2016/2/2b/T--HUST-China--notebook-PP-TT-fig01.png" alt="">
 
                         <img src="https://static.igem.org/mediawiki/2016/2/2b/T--HUST-China--notebook-PP-TT-fig01.png" alt="">
 
                     </p>
 
                     </p>
 +
                    <p>
 +
                        <h6>8.15</h6>
 +
                        <h6>DNA ligation&Transformation</h6>
 +
                        <h6>Ziyi Jiang</h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Backbone
 +
                                        <td>Strain
 +
                                            <td>Medium</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> PP2CA
 +
                                    <td>TTADH1-psb1c3(EcoRI and XbaI cut)
 +
                                        <td>DH5α
 +
                                            <td>LB(Chl added) </td>
 +
                            </tr>
 +
                                                  </table>
 +
                    </p>
 +
 
                     <p>
 
                     <p>
 
                         <h6>8.16</h6>
 
                         <h6>8.16</h6>
 
                         <h6>Colony PCR</h6>
 
                         <h6>Colony PCR</h6>
−
                         <h6>Successfully align the sequencing results with the objective sequences:</h6>
+
                         <h6>Successfully align the sequencing results with the objective sequences</h6>
 
                         <h6>Ziyi Jiang</h6>
 
                         <h6>Ziyi Jiang</h6>
 
                         <table border="1">
 
                         <table border="1">

Revision as of 06:42, 11 October 2016

Notebook

Notebook

see how these parts were constructed？


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
Colony PCR
Yufeng Mo
Number Name
1 CII 1
2 CII 2

CIII

7.14
PCR
Yufeng Mo
7.15
Restriction Enzyme Digestion
Yufeng Mo
Name Enzyme
plasmid psb1c3 1 XbaI and SpeI
plasmid psb1c3 2 XbaI and SpeI

7.16
Dephosphorylation
Yufeng Mo
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

RBS-CII

9.4
PCR
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

RBS-CIII

8.7
PCR
Ziyi Jiang
Name Total
RBS-CIII 50 μl

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

RBS-CIII-RBS-CIII

8.18
Restriction Enzyme Digestion
Ziyi Jiang
Name Enzyme
RBS-CIII EcoRI and SpeI
RBS-CIII EcoRI and XbaI

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

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 SpeI
RBS-CII-psb1c3 EcoRI and XbaI

9.14
DNA ligation&transformation
Yufeng Mo
Name Backbone Strain Medium
RBS-CII RBS-CII-psb1c3(EcoRI and XbaI 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

pRM-RBS-GFP-LAVtag

7.18
PCR
Xiaoyang Li
Name Total
GFP-F 50 μl

Name Total
pRM-RBS-GFP 1 30 μl
pRM-RBS-GFP 2 30 μl
pRM-RBS-GFP 3 30 μl

7.19
Restriction Enzyme Digestion
Name Enzyme
pRM-RBS-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

8.21
Restriction Enzyme Digestion
Yufeng Mo
Name Enzyme
pRM-RBS-GFP-LVAtag EcoRI and PstI
pETDuet-1 EcoRI and PstI

8.21
DNA ligation&transformation
Yufeng Mo
Name Backbone Strain Medium
pRM-RBS-GFP-LVAtag pETDuet-1(EcoRI 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

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

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

pR-RBS-GFP-LAVtag

8.2
PCR
Zhenyi Guo
Name Total
pR-RBS-GFP-tag 1 50 μl
pR-RBS-GFP-tag 2 50 μl
pR-RBS-GFP-tag 3 50 μl

8.2
Restriction Enzyme Digestion
Zhenyi Guo
Name Enzyme
psb1c3 1 EcoRI and PstI
psb1c3 2 EcoRI and PstI
pR-RBS-GFP-tag EcoRI and PstI

8.3
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
Zhenyi Guo
Successfully align the sequencing results with the objective sequences
Number Name
2 pR-RBS-GFP-tag

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(EcoRI 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

SnRK2.2

8.25
Site-mutated PCR
Fengyi Li
Name Total
SnRK2.2 40 μl

9.10
PCR
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

ABF2

9.10
PCR
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

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) DH5α LB(chl added)

7.27
Colony PCR
Yufeng Mo
Number Name
2 PP2CA

prd29A

9.27
PCR
Zhi Zeng
Name Total
prd29a 1 50 μl
prd29a 2 50 μl
prd29a 3 50 μl
prd29a 4 50 μl

9.27
Restriction Enzyme Digestion
Zhenyi Guo
Name Enzyme
prd29a 1 XbaI and PstI
prd29a 2 XbaI and PstI
9.27
DNA ligation&transformation
Zhenyi Guo
Name Backbone Strain Medium
prd29a psb1c3(XbaI and PstI cut) DH5α LB(chl added)

9.28
Colony PCR
Successfully align the sequencing results with the objective sequences
Zhenyi Guo

Number Name
1 prd29a-psb1c3
2 prd29a-psb1c3

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

8.17
DNA ligation&Transformation
Lin Di
Name Backbone Strain Medium
CI-TTADH1-pR-RBS-GFP-LVAtag Duet-pet(EcoRI and PstI cut) DH5α LB(Amp added)
CII-TTADH1-pRE-RBS-GFP-LVAtag Duet-pet(EcoRI and PstI cut) 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

Cro-TT-pRM-RBS-GFP-LVAtag-PSB1C3

8.14
PCR
Zhi Zeng
Name Total
Cro 80 μl

8.15
Restriction Enzyme Digestion
Zhi Zeng
Name Enzyme
pRM-RBS-GFP-LVAtag-psb1c3 EcoRI and XbaI
psb1c3 XbaI and PstI

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

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(EcoRI and PstI cut) 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

8.15
DNA ligation&Transformation
Ziyi Jiang
Name Backbone Strain Medium
PP2CA TTADH1-psb1c3(EcoRI and XbaI cut) DH5α LB(Chl added)

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

PP2CA-TTADH1-pCyc

8.22
Overlap PCR
Zhi Zeng
Name Total
PP2CA-TTADH1-Pcyc 80uL

8.22
Restriction Enzyme Digestion
Zhi Zeng
Backbone Enzyme
psb1c3 PstI and XbaI

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

Kozak-SnRK2.2-TTADH1-pCyc

8.30
Overlap PCR
Name Total
Kozak-Snrk2.2-TTADH1-pCyc 80uL

9.5
In-Fusion
Zhi Zeng
Insert Vector Total
Kozak-Snrk2.2-TTADH1-pCyc psb1c3(Cut by PstⅠ&XbaⅠ) 5uL

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

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

patp2-RBS-CI-TTADH1-pR-RBS-CIII

9.2
Overlap PCR
Jiyue Qin
Name Total
Patp2-RBS-CI-TT 50uL

9.3
In-Fusion
Jiyue Qin
Insert Vector Total
Patp2-RBS-CI-TT&pR-RBS-CIII psb1c3(Cut by PstⅠ&XbaⅠ) 10uL

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

RBS-iLDH-TT-pRM-RBS-beta-galactosidase

9.1
Restriction Enzyme Digestion
Jiyue Qin
Name Enzyme
GFP-LVAtag-psb1c3 XbaI and PstI

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

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

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