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