GSM5009765: V-D.gIL1RN.22nt-2; Homo sapiens; RNA-Seq
The RNA-seq analysis has been described in our previous work. Briefly, total RNA was isolate with TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and mRNA was enriched and fragmented. Deep-seq was used to determine the indel frequency. Briefly, total DNA was extracted using sarkosyl lysis buffer and on-target sites were amplified by PCR. RNA libraries were constructed by the following steps: the first strand synthesis, the second strand synthesis, adaptor adding, and PCR amplification. After verification and quantification, libraries were sequenced by HiSeq instrument with 150bp paired-end module. DNA libraries were constructed by the following steps: on-target sites were amplified by PCR with indexed forward and indexed reverse primers. After purification and concentration normalization, PCR products were pooled into different libraries. Completed libraries were generated by a second round of PCR using the pooled libraries as templates and sequenced with 150-bp paired-end reads on Illumina HiSeq instrument.