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Total RNA was isolated from different samples using RNeasy plant mini kits (TianGen, China) according to the manufacturer’s instructions and treated with RNase-free DNase I (Takara, Japan) to degrade genomic DNA. The quality and quantity of RNA was checked by Nanodrop 1000 Spectrophotometer (Thermo Fisher Scientific, Wilmington, DE) and Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). A total of 3 μg RNA per sample was used for cDNA library preparation. Sequencing libraries were constructed by NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, Ipswich, MA, USA) following the manufacturer’s instructions. Briefly, the steps included mRNA enrichment, mRNA fragmentation, second-strand cDNA synthesis, size selection, and PCR amplification were performed as previously described. The prepared libraries were then sequenced on an Illumina HiSeq 2000 platform (San Diego, CA, USA) |
RNA-Seq |
TRANSCRIPTOMIC |
PCR |
PAIRED
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