title: GSM8602149: Bulk RNA-seq, gRNA6, BE3, Day4, R1; Mus musculus; OTHER
library_construction_protocol: RNA was extracted using the RNeasy Micro Kit (Qiagen, 74004) for bulk barcoding at various time points after cell sorting. To amplify the gRNA loci, reverse transcription was performed with 1 µl of a primer and 500 ng of RNA. cDNA was amplified in a 20 µl reaction using KAPA HiFi HotStart ReadyMix (KAPA Biosystems, KK2502) with specific primers. PCR products were purified using AMPure XP Beads and sequenced on a single end Illumina MiSeq run with 58 cycles and an 8-cycle index.