GSM8787963: HEK293T_SRRM2_RTTag_n5_(240518,241018_NK_hs_2157); Homo sapiens; OTHER
The detailed RT&Tag protocol can be found here: dx.doi.org/10.17504/protocols.io.x54v9jyjqg3e/v1. Sequencing libraries were then amplified by mixing the nuclei suspension with 15 μL of 0.67% Triton X-100, 2 μL of 10 mM i7 primer, 2 μL of 10 mM i5 primer, and 25 μL of 2x NEBNext Master Mix (NEB). The following PCR conditions were used: 1) 58°C for 5 minutes, 2) 72°C for 5 minutes, 3) 98°C for 30 seconds, 4) 98°C for 10 seconds, 5) 60°C for 15 seconds, 6) Repeat steps 4-5 13 times, 7) 72°C for 2 minutes, 8) Hold at 4°C. The sequencing libraries were then purified using 0.8x HighPrep PCR Cleanup System (MagBio) beads according to the manufacturer's instructions. Sequencing was performed using single-end 100 bp reads.