title: Illumina MiSeq paired end sequencing; MiSeq 2x300 paired-end sequencing of HEK293T cells stably expressing eGFP-PPP2R5A transduced with a HIV-1 Vif mutant library
design: MiSeq 2x300 paired-end sequencing of HEK293T cells stably expressing eGFP-PPP2R5A transduced with a HIV-1 Vif mutant library
library_construction_protocol: Cells were sorted using fluorescence-activated cell sorting, pelleted, and frozen.  Nucleic acids were extracted from frozen cell pellets using a Qiagen all prep DNA/RNA extraction kit One microgram of purified RNA was reverse-transcribed and PCR amplified using gene specific primers with truncated Illumina adapter primers. The initial PCR reaction was separated on a DNA agarose gel and the PCR band was extracted and gel purified using a DNA extraction kit (Thermo). The purified product was amplified using the final segments of the Illumina adapter primers and purified using a DNA clean-up kit (Thermo). Samples were then quantified and submitted for Illumina MiSeq analysis.