| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
Total RNA was extracted using CTAB method, and then subsequently treated with RNase-Free DNase I (Qiagen, USA) to eliminate genomic DNA. The RNA integrity number (RIN) of each sample was determined using an Agilent 2100 Bioanalyzer, and samples with an integrity number >8.5 were used for library construction. mRNA was enriched from 20 μg of total RNA using oligo (dT) magnetic beads,and then cleaved into small pieces using fragmentation buffer. The first cDNA strand was synthesized using random hexamer primers and reverse transcriptase, followed by second-strand cDNA synthesis using DNA polymerase I and RNase H. The short double-stranded fragments were purified with a QiaQuick PCR Purification Kit (Qiagen, CA, USA), and dissolved with the EB buffer supplied in the kit for end preparation and poly(A) addition. The sequence adaptors were linked to two ends of the short cDNA sequences. Agarose gel electrophoresis was used to select suitably sized fragments, and the products were subsequently amplified by PCR. |
RNA-Seq |
TRANSCRIPTOMIC |
unspecified |
PAIRED
|
|
