What is vector recombination, and how do I prevent it?
Vectors have a natural tendency to produce background recombinants, by recombining at the long terminal repeats (LTR’s). If this occurs, it will result in loss of most of the plasmid and can confer a strong growth advantage on the cells. It is therefore critical to maintain rigid growth conditions to minimize the abundance of background recombinants. This includes: i. Avoiding extended incubation times, longer than 12-16 hours; ii. Avoiding repeated freeze-thaw cycles; iii.Always work from fresh, single-colony selections; iv.Growth of clones in low-salt (5g NaCl/L) LB media (Lennox); Open Biosystems has also designed the pGIPZ vector to ensure high quality and minimal recombination; with the above growth conditions maintained, the pGIPZ vector will display negligible background recombination. Minimal levels of recombination have been verified by Open Biosystems through extensive, repeated internal QC processes: Representative shRNAmir containing pGIPZ lentiviral clones grown for 16 hou
