Cloning of Solute Carriers (SLCs) into pHTBV1.1 Bacmid for High-Throughput Protein Expression

To begin, add 150 nanograms of solute-carrier, or SLC clone, and 100 nanograms of the vector in a 96-well plate. Make the reaction volume to 8 microliters with 10 millimolar tris solution pH 8. Then add 2 microliters of recombinant enzyme mix and incubate for one hour at room temperature.

After incubation, add 1 microliter of proteinase K and incubate for 30 minutes at 37 degrees Celsius. Add 4 microliters of reaction mixture to 50 microliters of chemically competent E.coli Mach1 cells, and transform the cells using the heat shock method. After transformation, plate the bacterial suspension onto an LB agar plate.

Identify colonies harboring the pHTBV1.1 vector with the SLC gene insert using appropriate primers and standard protocols for colony PCR.