GSM4192671: R1T1_200ms_lib; Homo sapiens; OTHER
After sorting, cells in each Dendra2 photoactivation bin were grown in the absence of doxycycline until confluent in one well of a 6-well plate (~ 7 days), then pelleted and stored at -20 °C.  DNA was extracted from cell pellets with the DNEasy kit (Qiagen 69504) using RNAse according to the manufacturer's instructions. gDNA was amplified using SV40_NLS_seq_f and SV40_NLS_seq_r (Supplementary Table 8) primers using Kapa Hifi (Kapa Biosystems KK2602) according to the manufacturer's instructions. Amplicons were cleaned using Ampure XP beads (Beckman Coulter A63880), then subjected to an indexing PCR step using KAPA2G Robust (Kapa Biosystems KK5705) with primers P5 and an indexing primer. Amplicons were then run on a 1.5% agarose gel at 130 V for 40 min and the DNA in the 235bp band extracted using Freeze'N Squeeze DNA Gel Extraction Spin Columns (BioRad 7326165). Extracted DNA was sequenced on an Illumina NextSeq500 using SV40_NLS_Read1, SV40_NLS_Read2, and SV40_NLS_Index1 primers.
GEO Accession
GSM4192671