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| 文库名称 |
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Total RNA was extracted using Trizol reagent kit
according to the manufacturer’s protocol. RNA quality was assessed on an Agilent 2100
Bioanalyzer and checked using RNase
free agarose gel electrophoresis. After total RNA was extracted, mRNA was
enriched by Oligo(dT) beads, Then the enriched
mRNA was fragmented into short fragments using fragmentation buffer and reverse
transcribed into cDNA with random primers. Second-strand cDNA was synthesized by
DNA polymerase I, RNase H, dNTP and buffer. Then the cDNA fragments were
purified with QiaQuick PCR extraction kit, end
repaired, poly(A) added, and ligated to Illumina sequencing adapters. The ligation
products were size selected by agarose gel electrophoresis, PCR amplified, and
sequenced using Illumina HiSeq
TM
4000 |
RNA-Seq |
TRANSCRIPTOMIC |
PolyA |
PAIRED
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