Forward genetic screen for mutagenic human transposons using HPRT1 gene trapping
The human HPRT1 gene was amplified by long-range PCR using amplicon primers and conditions as described in the Supplementary Methods. Equimolar amounts of purified amplicons were pooled according to concentration provided by Qubit (Invitrogen Carlsbad, CA) and sized using the 2100 BioAanalyzer instrument (Agilent Technologies, Santa Clara, CA). The sequencing library construction was performed using the KAPA Hyper Prep kit (#KK8504, KAPA biosystems, Wilmington, MA) and indexed Illumina adaptors from IDT (Coralville, IO), strictly following manufacturer’s instructions. After quantification and sizing, libraries were pooled for sequencing on using Miseq (pooled library input at 10pM) with 300/300 paired-end sequencing.