GSM2790962: 18S rRNA 1045+1056_CMC-; Homo sapiens; RNA-Seq
Total RNA was extracted, fragmented and then CMC-reacted or mock-treated, followed by developed reverse transcription using our method. The RT-PCR products of specific qPCR amplicons were subjected to high-throughput sequencing. Libraries were prepared using NEBNext® Ultra II DNA Library Prep Kit. Briefly, DNA was end-repaired by an enzyme mix to yield a protruding 3'- 'A' base for ligation with our in-house synthesized adaptors which have a single 'T' base overhang at the 3’ end. After adaptor ligation, DNA was PCR amplified with Illumina primers for 15 cycles, followed by size-selection and purification using AMPure XP beads (New England Biolabs). Libraries were sequenced on Illumina HiSeq X Ten with double end reads (150 bp).