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The RNA was converted into Illumina-compatible sequencing libraries with MINERVA protocol. Briefly, five miul of nucleic acid was digested with DNase I(New England Biolabs, US), and then the first-strand cDNA was synthesized with Oligo dT primer(5'-T30VN-3'), random decamer primer (5'-N10-3'), and SuperScript II reverse transcriptase (Invitrogen, Carlsbad, CA). Direct tagmentation of RNA/DNA hybrids was using Tn5 transposase (Vazyme, Nanjing, CHN), followed by index PCR to amplify the library. Finally, the product was purified with AMPure XP beads (Beckman Coulter, MA, US). All purified libraries were sequenced on Illumina NovaSeq with 2x150 paired-end mode. |
RNA-Seq |
METATRANSCRIPTOMIC |
unspecified |
PAIRED
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