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The first strand of cDNA was synthesized in the M-MuLV reverse transcriptase system using fragmented mRNA as template and random oligonucleotides as primers, followed by RNaseH degradation of the RNA strand, and the second strand of cDNA was synthesized with dNTPs as raw material under DNA polymerase I system. The purified double-stranded cDNA underwent end repair, A-tail, and sequencing adapter ligation, and the cDNA of about 250-300 bp was screened with AMPure XP beads, PCR amplification was performed, and the PCR product was purified again using AMPure XP beads to obtain the library. |
RNA-Seq |
TRANSCRIPTOMIC |
PCR |
PAIRED
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