GSM7912296: c3; Mus musculus; RNA-Seq
RNA was harvested using Takara TRIzol according to manufacturer's instruction. Then total RNA was treated by either mRNA enrichment or rRNA removal. The obtained RNA was segmented by interrupting buffer, and then reverse-transcribed by random N6 primers, and then synthesized cDNA double strand to form double-stranded DNA. The end of the synthetic double-stranded DNA is patched and phosphorylated at the 5 'end, forming A sticky end with a protruding "A" at the 3' end, and then connected to a bubbling splice with a protruding "T" at the 3' end. The linked products were amplified by PCR using specific primers. The PCR products are thermologically denatured into single strand, and then the single strand DNA is cycled with a bridge primer to obtain a single strand circular DNA library.