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Past Tasks - Revision history
2026-10-18T04:56:44Z
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Paul.xu: Created page with "== Sam: Protein Purification == '''Next Week''' 1. Cre recombinase 1708 pT7A3-HTNCre 2. 1697 pCold-GST-(3c)-wt3C or 1613 pET28-cold-GST-3C 3. Screening of Protei..."
2017-10-10T21:06:01Z
<p>Created page with "== Sam: Protein Purification == '''Next Week''' 1. Cre recombinase 1708 pT7A3-HTNCre 2. 1697 pCold-GST-(3c)-wt3C or 1613 pET28-cold-GST-3C 3. Screening of Protei..."</p>
<p><b>New page</b></p><div>== Sam: Protein Purification ==<br />
<br />
'''Next Week'''<br />
<br />
1. Cre recombinase 1708 pT7A3-HTNCre<br />
<br />
2.&nbsp;1697 pCold-GST-(3c)-wt3C or&nbsp;1613 pET28-cold-GST-3C<br />
<br />
3. Screening of Protein Expression:<br />
<br />
820 pHis-GST-HA-His<br />
<br />
819 pHis-GST-myc-His<br />
<br />
818 pHis-GST-His-Flag-His<br />
<br />
1697 pCold-GST-(3c)-wt3C<br />
<br />
1613 pET28-cold-GST-3C<br />
<br />
2462 pT7A3E-Tn5-ChiBD-6H<br />
<br />
== Daniel: Cloning & Expression Screening ==<br />
<br />
832 pMiniT-VVTopo<br />
<br />
823 pMiniT-T5Exo<br />
<br />
<br />
<br />
827 pHis-MBP-HA-His<br />
<br />
826 pHis-MBP-myc-His<br />
<br />
825 pHis-MBP-His-Flag-His<br />
<br />
Use 1661 pT7A3-MBP-(3c)-NRF2 as the source of MBP, so you need to grow up pT7A3-MBP-(3c)-NRF2 and make minipep ASAP<br />
<br />
824 pCold-GST-VVTopo<br />
<br />
2443 pET28b-Tn5<br />
<br />
2474 pET30-HA-Tn5-6His<br />
<br />
'''Next Week'''<br />
<br />
== Paul ==<br />
<br />
1. Purification of 1032 pET6XHis-sso7d-wt-Taq sso7d-Taq<br />
<br />
2. Clone 2474 pET30-HA-Tn5-6His<br />
<br />
3, Clone 2443 pET28b-Tn5TPnase<br />
<br />
4. QC Taq for qPCR 6. Transfection QC<br />
<br />
5. Express&nbsp;1881 pET30b-wt-Xho1 +&nbsp;1878 pLysN-Plac-mXho1<br />
<br />
6, Clone&nbsp;<br />
<br />
808 1032 pET6XHis-sso7d-EP-Taq<br />
<br />
810 1032 pET-FastTaq-A743H<br />
<br />
809 1032 pET-FastTaq-E742A<br />
<br />
2495 pMiniT-TP4-kLC-vr<br />
<br />
2494 pMiniT-TP4-HC-vr<br />
<br />
2493 pCMV5-mIgG2a-kLC<br />
<br />
2492 pCMV5-mIgG2a-HC<br />
<br />
2491 pMiniT-mIgG2a-kLC<br />
<br />
2490 pMiniT-mIgG2a-HC<br />
<br />
820 pHis-GST-HA-His<br/>819 pHis-GST-myc-His<br/>818 pHis-GST-His-Flag-His<br/>821 pCold-GST-(3C)-T5Exo<br />
<br />
'''Design'''&nbsp;thermostable luciferase,<br />
<br />
Antibody Fusion &nbsp;vector for IgG2a from 1162 pFUSE2-CLIg-hk 1161 pFUSE-CHIg-hG1 both of them from 1101<br />
<br />
Protein A,<br />
<br />
new bright GFP,<br />
<br />
Dre<br />
<br />
Cre-Flp<br />
<br />
PD-1<br />
<br />
PDL-1<br />
<br />
Cas9<br />
<br />
Cid1<br />
<br />
T5-Exo<br />
<br />
etc<br />
<br />
Taq Polymerase primer-dimer QC<br />
<br />
freeze dry new Taq, chemical modification, QC<br />
<br />
design c-Myc 9E10 bacterial production plasmids<br />
<br />
<span style="color: rgb(255, 0, 0);">'''Purify important enzymes: for Sam'''</span><br />
<br />
483 pET28b-TthPol (for RT-qPCR)<br />
<br />
1995 Kfu<br />
<br />
1811 VPol<br />
<br />
1458 KOD<br />
<br />
1483 T4 DNA Ligase<br />
<br />
1464 T7 DNA Ligase<br />
<br />
2409 pT7A3-Tat-dCre-(Tev)-Tat-NLS-iFLP-6H<br />
<br />
1567 pT7A3-T5-Exo<br />
<br />
1715 pT7A30-Pfu-sso7d pI is 8.47 / 98.7<br />
<br />
1765&nbsp;pT7A3-T4DNAPol &nbsp; 6.19 / 105.9<br />
<br />
1722 pT7A3-sso7d-Native-ToughTaq(E708K)<br />
<br />
1542 pT7A3-sso7d-Native-ToughTaq(E708K)<br />
<br />
1497 &nbsp; pT7A3-ToughTaq(gene synthesis)<br />
<br />
<span style="color: rgb(255, 0, 0);">Purify important enzymes: for Paul</span><br />
<br />
<br />
<br />
Transform competent cells with 1002 pLysN<br />
<br />
DNA ladder<br />
<br />
10X100 bp, 750 bp, 1500 bp, 2500 bp<br />
<br />
4X250bp, 2X500 bp, 1 x 1000 bp<br />
<br />
GR Green GR Red Stain to make dual color ladder<br />
<br />
GR Green stain ladder from Ebay, GRRed stain of PCR product<br />
<br />
ppt using PEG, reconsititute in 6X loading dye<br />
<br />
make pMiniT vector<br />
<br />
poly (U) polymerase 2252 pT7A3-Cid1,<br />
<br />
gene synthesis shrimp alkaline phosphatase (SAP)<br />
<br />
calf intestinal phosphatase (CIP)<br />
<br />
bovine alkaline phosphatase (BAP)<br />
<br />
&nbsp;<br />
[[Category:Tasks]]</div>
Paul.xu