GSM9111439: SW1 sh Polr1a biol rep 1; Mus musculus; RNA-Seq
RNA was isolated using the Rneasy kit (Qiagen). Magnetic beads with Oligo (dT) were used to isolate mRNA from the total RNA. The mRNA was fragmented randomly by adding the fragmentation buffer, then the cDNA was synthesized by using mRNA template and random hexamers primer after adding dNTPs, RNase H, and DNA polymerase I. Short fragments were purified and resolved with EB buffer for end repair and single-nucleotide A (adenine) addition. After that, the short fragments were connected with sequencing adapters. The double-stranded cDNA library was completed through size selection and PCR enrichment. During the QC steps, the Agilent 2100 Bioanalyzer and ABI StepOnePlus Real-Time PCR System was used in quantification and qualification of the sample library. Lastly, the qualified RNA-seq libraries are sequenced using Illumina NovaSeq6000.