Past Tasks

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 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

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

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)