E coli max efficiency stbl2

Infectious molecular clone NL4-3 was obtained from the NIH AIDS Reagent Program. BocLys was purchased from Bachem. Primers were ordered from Sigma. Restriction enzymes and T4 DNA ligase were purchased from New England Biolabs. KOD hot start DNA polymerase was purchased from EMD Millipore. Standard molecular biology techniques³² were used throughout. Site-directed mutagenesis was carried out using overlapping PCR. E. coli GeneHogs were used for routine cloning and DNA propagation for none-HIV plasmids. E. coli MAX Efficiency Stbl2 (Thermo Fisher Scientific Inc) was used for routine cloning and DNA propagation for HIV plasmids. All solutions were prepared in deionized water further treated by Barnstead Nanopure® ultrapure water purification system (Thermo Fisher Scientific Inc). Antibiotics were added where appropriate to following final concentrations: ampicillin, 100 mg L^-1; kanamycin, 50 mg L^-1; tetracycline, 12.5 mg L^-1.

AzF was purchased from Bachem. Primers were ordered from Sigma. Restriction enzymes, Antarctic phosphatase (AP) and T4 DNA ligase were purchased from New England Biolabs. KOD hot start DNA polymerase was purchased from EMD Millipore. Standard molecular biology techniques³⁶ were used throughout. Site-directed mutagenesis was carried out using overlapping PCR. E. coli GeneHogs were used for routine cloning and DNA propagation for none-HIV plasmids. E. coli MAX Efficiency Stbl2 (Thermo Fisher Scientific Inc) was used for routine cloning and DNA propagation for HIV plasmids. All solutions were prepared in deionized water further treated by Barnstead Nanopure® ultrapure water purification system (Thermo Fisher Scientific Inc). Antibiotics were added where appropriate to following final concentrations: ampicillin, 100 mg L⁻¹; kanamycin, 50 mg L⁻¹; tetracycline, 12.5 mg L⁻¹.