| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
Total RNA was extracted using TRIZOL reagent (Takara Bio Inc., Otsu, Japan). The degradation and purity of total RNA were examined through 1% agarose electrophoresis analysis and the NanoPhotometer spectrophotometer (IMPLEN, CA, USA), respectively. The integrity was also further demonstrated with the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). The cDNA library was constructed and sequenced by Novogene (Beijing, China). In brief, poly-Toligo-attached magnetic bead was used to purify the mRNA from total RNA which was subsequently fragmented with divalent cations under enhanced temperature. Then, the fragmented mRNA was used as the template for the synthesis of the first strand of cDNA with M-MuLV Reverse Transcriptase and a random hexamer primer. The templates were degraded by RNaseH followed by mixing with DNA polymerase I, and dNTPs to synthesize the second strand of cDNA. After purification with the AMPure XP system (Beckman Coulter, Beverly, USA), the double-stand DNA was enriched and amplified by PCR. After being purified using the AMPure XP system, the library quality was analyzed on the Agilent Bioanalyzer 2100 system and sequenced on an Illumina Hiseq platform according to the manufacturer’s instructions. |
RNA-Seq |
TRANSCRIPTOMIC |
cDNA |
SINGLE
|
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