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

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                     </p>
 
                     </p>
 
                 </div>
 
                 </div>
 +
<div>
 +
                    <h2 class="title"> PRE-RBS-Cro-RBS-CII-TT-ptrp </h2>
 +
                    <p>
 +
                        <h6>10.1</h6>
 +
                        <h6>PCR</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Total</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> PRE-RBS-Cro-RBS-CII-TT-ptrp
 +
                                    <td>50 μl</td>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.2</h6>
 +
                        <h6>Restriction Enzyme Digestion</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Enzyme</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> Psb1c3
 +
                                    <td> XbalI and PstI </td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
src="https://static.igem.org/mediawiki/2016/3/3d/T--HUST-China--GZY-2.jpeg
 +
"">
 +
</div>
 +
 +
                        <h6>10.3</h6>
 +
                        <h6>DNA ligation&transformation</h6>
 +
                        <h6> Zhenyi Guo  </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name</td>
 +
                                <td>Backbone</td>
 +
                                <td>Strain</td>
 +
                                <td>Medium</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> PRE-RBS-Cro-RBS-CII-TT-ptrp </td>
 +
                                <td>psb1c3</td>
 +
                                <td>DH5α</td>
 +
                                <td>LB(chl added) </td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
 +
 +
                        <h6>10.4</h6>
 +
                        <h6>Colony PCR</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Number</td>
 +
                                <td>Name</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td>1</td>
 +
                                <td> PRE-RBS-Cro-RBS-CII-TT-ptrp- PSB1C3</td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <img src="https://static.igem.org/mediawiki/2016/6/65/T--HUST-China--GZY-4.jpeg
 +
"">
 +
</div>               
 +
<div>
 +
                  <h2 class="title"> RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag </h2>
 +
                    <p>
 +
                        <h6>10.1</h6>
 +
                        <h6>PCR</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Total</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag                                    <td>50 μl</td>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.2</h6>
 +
                        <h6>Restriction Enzyme Digestion</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Enzyme</td>
 +
                            </tr>
 +
                          <tr>
 +
  <td>Psb1c3</td>
 +
  <td>XbalI and PstI</td>
 +
</tr>
 +
<tr>
 +
  <td>RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag</td>
 +
  <td>XbalI and PstI</td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
<img src=" https://static.igem.org/mediawiki/2016/8/89/T--HUST-China--GZY-3.jpeg
 +
”">
 +
 +
                        <h6>10.3</h6>
 +
                        <h6>DNA ligation&transformation</h6>
 +
                        <h6> Zhenyi Guo  </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name</td>
 +
                                <td>Backbone</td>
 +
                                <td>Strain</td>
 +
                                <td>Medium</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag </td>
 +
                                  <td>psb1c3</td>
 +
                                <td>DH5α</td>
 +
                                <td>LB(chl added) </td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.4</h6>
 +
                        <h6>Colony PCR</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Number</td>
 +
                                <td>Name</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td>1</td>
 +
                                <td> RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag - Ppsb1c3</td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <img src=" https://static.igem.org/mediawiki/2016/7/7f/T--HUST-China--GZY-5.jpeg"">
 +
                </div>
 +
<div>
 +
                    <h2 class="title"> PRE-RBS-Cro-RBS-CII-TT-ptrp </h2>
 +
                    <p>
 +
                        <h6>10.1</h6>
 +
                        <h6>PCR</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Total</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> Kozak-ABF2-TTADH1-pRD29A-Kozak-GFP-LVAtag
 +
                                    <td>50 μl</td>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.2</h6>
 +
                        <h6>Restriction Enzyme Digestion</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Enzyme</td>
 +
                            </tr>
 +
                            <td>pCyc-psb1c3</td>
 +
  <td>EcoRI and XbalI</td>
 +
</tr>
 +
<tr>
 +
  <td>Kozak-ABF2-TTADH1-pRD29A-Kozak-GFP-LVAtag</td>
 +
  <td>XbalI and PstI</td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.3</h6>
 +
                        <h6>DNA ligation&transformation</h6>
 +
                        <h6> Zhenyi Guo  </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name</td>
 +
                                <td>Backbone</td>
 +
                                <td>Strain</td>
 +
                                <td>Medium</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> Kozak-ABF2-TTADH1-pRD29A-Kozak-GFP-LVAtag </td>
 +
                                <td>psb1c3</td>
 +
                                <td>DH5α</td>
 +
                                <td>LB(chl added) </td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.4</h6>
 +
                        <h6>Colony PCR</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Number</td>
 +
                                <td>Name</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td>1</td>
 +
                                <td> pCyc-Kozak-ABF2-TTADH1-pRD29A-Kozak-GFP-LVAtag -psb1c3</td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    </div>
 +
<div>
 +
                    <h2 class="title"> RBS-ompA-Ildh-TT-pRM-RBS-beta-galactosidase-Placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII </h2>
 +
                    <p>
 +
                        <h6>10.6</h6>
 +
                        <h6>PCR</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Total</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> RBS-ompA-iLDH                                    <td>50 μl</td>
 +
</tr>
 +
<tr>
 +
  <td>TT-pRM-RBS-beta-galactosidase</td>
 +
  <td>50μl</td>
 +
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.7</h6>
 +
                        <h6>Restriction Enzyme Digestion</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name
 +
                                    <td>Enzyme</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> Placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-psb1c3                                    <td> EcoRI and XbalI </td>
 +
            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.8</h6>
 +
                        <h6>DNA ligation&transformation</h6>
 +
                        <h6> Zhenyi Guo  </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Name</td>
 +
                                <td>Backbone</td>
 +
                                <td>Strain</td>
 +
                                <td>Medium</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td> RBS-ompA-iLDH、TT-pRM-RBS-beta-galactosidase </td>
 +
                                <td> Placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-psb1c3</td>
 +
                                <td>DH5α</td>
 +
                                <td>LB(chl added) </td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <p>
 +
                        <h6>10.9</h6>
 +
                        <h6>Colony PCR</h6>
 +
                        <h6> Zhenyi Guo </h6>
 +
                        <table border="1">
 +
                            <tr>
 +
                                <td>Number</td>
 +
                                <td>Name</td>
 +
                            </tr>
 +
                            <tr>
 +
                                <td>1</td>
 +
                                <td> RBS-ompA-Ildh-TT-pRM-RBS-beta-galactosidase-Placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-psb1c3</td>
 +
                            </tr>
 +
                        </table>
 +
                    </p>
 +
                    <img src="https://static.igem.org/mediawiki/2016/d/d8/T--HUST-China--GZY-1.jpeg
 +
"">
 +
</div>
 +
 
             </section>
 
             </section>
 
         </div>
 
         </div>

Revision as of 17:08, 19 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

CIII

7.14
PCR
Yufeng Mo
Name Total
CIII 1 50 μl
7.15
Restriction Enzyme Digestion
Yufeng Mo
Name Enzyme
plasmid psb1c3 1 XbaI and SpeI
CIII XbaI and SpeI

7.15
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
Colony PCR
Yufeng Mo
Number Name
1 CIII 1
2 CIII 2
3 CIII 3
4 CIII 4
5 CIII 5
6 CIII 6

RBS-CII

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
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
Ziyi Jiang
Name Backbone Strain Medium
RBS-CIII psb1c3(XbaI and PstI cut) DH5α LB(chl added)

8.11
Colony PCR
Ziyi Jiang
Number Name
2 RBS-CIII

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
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-psb1c3 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 XbaI cut) DH5α LB(chl added)

9.15
Colony PCR
Zhi Zeng
Number Name
1 RBS-CII-RBS-CII-psb1c3

pRM-RBS-GFP-LAVtag

7.18
PCR
Xiaoyang Li
Name Total
GFP-First 50 μl

Name Total
pRM-RBS-GFP-LAVtag 30 μl
pRM-RBS-GFP-LAVtag 30 μl
pRM-RBS-GFP-LAVtag 30 μl

7.19
Restriction Enzyme Digestion
Xiaoyang Li
Name Enzyme
GFP EcoRI and SpeI
psb1c3 EcoRI and SpeI

7.20
DNA ligation&transformation
Xiaoyang Li
Name Backbone Strain Medium
pRM-RBS-GFP-LAVtag 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-pETDuet 1
3 pRM-RBS-GFP-LVAtag-pETDuet 3
4 pRM-RBS-GFP-LVAtag-pETDuet 4
5 pRM-RBS-GFP-LVAtag-pETDuet 5
6 pRM-RBS-GFP-LVAtag-pETDuet 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 50 μl
pR-RBS-GFP-tag 50 μl
pR-RBS-GFP-tag 50 μl

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

8.2
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
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

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
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) E.coli 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 XbaI and PstI

9.28
Colony PCR
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
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

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 (ligated by In-fusion) 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
PETDuet-1 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 pETDuet-1(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
DNA ligation&Transformation
Ziyi Jiang

8.16
Colony PCR
Ziyi Jiang
Name Backbone Strain Medium
PP2CA TTADH1-psb1c3(EcoRI and XbaI cut) DH5α LB(Chl added)
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 (ligated 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(ligated 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(ligated 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 1 50uL
Patp2-RBS-CI-TT 2 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
pETDuet-1 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

9.17
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 pETDuet-1(cut by EcoRI and PstI) DH5α LB(Amp added)

9.18
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-Duet-pet

PRE-RBS-Cro-RBS-CII-TT-ptrp

10.1
PCR
Zhenyi Guo
Name Total
PRE-RBS-Cro-RBS-CII-TT-ptrp 50 μl

10.2
Restriction Enzyme Digestion
Zhenyi Guo
Name Enzyme
Psb1c3 XbalI and PstI

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10.3
DNA ligation&transformation
Zhenyi Guo
Name Backbone Strain Medium
PRE-RBS-Cro-RBS-CII-TT-ptrp psb1c3 DH5α LB(chl added)

10.4
Colony PCR
Zhenyi Guo
Number Name
1 PRE-RBS-Cro-RBS-CII-TT-ptrp- PSB1C3

RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag

10.1
PCR
Zhenyi Guo
Name Total
RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag 50 μl

10.2
Restriction Enzyme Digestion
Zhenyi Guo
Name Enzyme
Psb1c3 XbalI and PstI
RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag XbalI and PstI

10.3
DNA ligation&transformation
Zhenyi Guo
Name Backbone Strain Medium
RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag psb1c3 DH5α LB(chl added)

10.4
Colony PCR
Zhenyi Guo
Number Name
1 RBS-CI-TT-pR-RBS-CIII-RBS-RFPtag-TT-pRM-RBS-GFPtag - Ppsb1c3

PRE-RBS-Cro-RBS-CII-TT-ptrp

10.1
PCR
Zhenyi Guo
Name Total
Kozak-ABF2-TTADH1-pRD29A-Kozak-GFP-LVAtag 50 μl

10.2
Restriction Enzyme Digestion
Zhenyi Guo
Name Enzyme
pCyc-psb1c3 EcoRI and XbalI
Kozak-ABF2-TTADH1-pRD29A-Kozak-GFP-LVAtag XbalI and PstI

10.3
DNA ligation&transformation
Zhenyi Guo
Name Backbone Strain Medium
Kozak-ABF2-TTADH1-pRD29A-Kozak-GFP-LVAtag psb1c3 DH5α LB(chl added)

10.4
Colony PCR
Zhenyi Guo
Number Name
1 pCyc-Kozak-ABF2-TTADH1-pRD29A-Kozak-GFP-LVAtag -psb1c3

RBS-ompA-Ildh-TT-pRM-RBS-beta-galactosidase-Placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII

10.6
PCR
Zhenyi Guo
Name Total
RBS-ompA-iLDH 50 μl
TT-pRM-RBS-beta-galactosidase 50μl

10.7
Restriction Enzyme Digestion
Zhenyi Guo
Name Enzyme
Placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-psb1c3 EcoRI and XbalI

10.8
DNA ligation&transformation
Zhenyi Guo
Name Backbone Strain Medium
RBS-ompA-iLDH、TT-pRM-RBS-beta-galactosidase Placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-psb1c3 DH5α LB(chl added)

10.9
Colony PCR
Zhenyi Guo
Number Name
1 RBS-ompA-Ildh-TT-pRM-RBS-beta-galactosidase-Placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-psb1c3