Shanghai 吉玛 System 药 technology 吉玛重组慢病 Poison Manual
Contents
1. tip 70 0 6 1h Lentivirus GenePharma Lentivirus VSVG Lentivirus2. GFP Lentivirus 72 96h GFP GFP Lentivirus MOI 4hrs 8hrs 12hrs 10 10 10 10 10 10 Lentivirus ATCC No 80 MOI 293T CRL 11268 lt 1 95D
3. TU ml 2 5 10 3 pot blot RNA dot blot RNA ml DNA TU ml RNA DNA 1 1000 2 1 3 5X103 96 100uL 30 50 2 MOI Multiply of Infection 5ug mL Polybrene Polybrene 3 90uL 4 2
4. 1 Fig 1 2 293T 1 5 3 BT TU ml transducing units TU nl PxN 100 x V x1 DF P GFP cells N V n1 DF dilution factor 1 undiluted 10 1 diluted 1 10 10 2 diluted 1 100 GFP p24 ELISA Kit ng p24 ml 102 103 LP ml 3 TU ml RNA RT qPCR RNA ml 102 104 Vg ml 2 TU ml GFP FACS TU ml
5. 10 3 Lentivirus Polybrene MOI Polybrene Polybrene Lentivirus 293T BHK21 24hrs GFP GFP 72 96hrs GFP GFP Lentivirus 80C
6. gt 100 0 5 021 51320195 51370738 51370739 021 51320295 1011 201203 www genepharma com E MAIL service genepharma com 218 A3 0512 6295 6678 0512 6295 6679 www genepharma com
7. 80C 6 6 12 ES 80 C 37 C 4 C 1 HIV 4kb 2 Lentivirus MOI Polybrene 4 8hg ml 3 Lentivirus GFP GFP
8. Lentivirus MOI in vivo 1 1 293T 96 0 5 10 X10 100ul 2 7 10 EP 90ul DMEM 10 FBS 3 10ul 10ul 4 90ul 37 C 5 CO2 5 48 100ul 6 3 4
9. EP 10ul 1X108TU ml 80C 10ul 10 100 5 10uL MOI 100 10 1 1X108TU ml MOI 6 7 8 12 24 8 72 96 9
10. RSV enhancer promoter Tat 3 LTR U3 self inactivating SIN 3 3 Gag Pol Rev VSV G HIV 1 Env 3 pGag Pol pRev PpVSVG RCL 4 MuLV 5 4 HIV 1 MuLV 6 LTR 2 RCL BSL2
11. GenePharma ShangHai GenePharma Co Ltd GenePharma Recombinant Lentivirus Operation Manual ShangHai GenePharma Co Ltd 1 NC transfer vector RNA shRNA 1 1 HIV 1 HIV 1 2 3 LTR 5 LTR U3
12. 3 CNE 1 gt 50 hADSC 2 100 HEC 1A HTB 113 5 HepG 2 20 hFOB1 19 2 HL 60 CCL 240 50 HLE B3 1 hMSC 50 hSMC 50 HUVEC 5 Jurkat TIB 152 100 MCF 7 HTB 22 5 MDA MB 231 HTB 26 10 SaoS 2 20 SGC 7901 5 SHG 44 10 SKOV 3 HTB 77 3 SMMC 7721 1 SPC A 1 10 THP 1 TIB 202 100 U251 1 U 20S HTB 96 5 U937 CRL 1593 2 50 HSC T6 3 PC 3 CRL 1721 100 A549 H9c2 2 1 CRL 1446 1 KM3 er B 50 80 MOI 50 5
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