alias: Dietzia sp.111N12-1
title: Dietzia sp.111N12-1 transcriptomic
design: The cDNA library for Illumina sequencing was prepared following the instruction of the TruseqTM RNA sample prep Kit (Illumina). First, mRNA was isolated from the total quantified RNA using poly-T oligo-attached magnetic beads (Illumina). The mRNA was subsequently disrupted into fragments (200-700 nt) using divalent cations. First-strand cDNAs were then synthesized using SuperScript II reverse transcriptase with small-fragment RNAs as templates and second-strand cDNA synthesis by DNA polymerase I. Next, these cDNA fragments were end-repaired and 3’-adenylated using T4 DNA polymerase, Klenow DNA polymerase, and T4 polynucleotide kinase. Illumina’s paired-end adaptors were ligated to the ends of these 3’-adenylated cDNA fragments. The cDNA fragments with adaptors were enriched by 15 cycles of PCR amplification with Phusion DNA polymerase, and the products were electrophoresed on a 2%-agarose gel (certified low-range, ultra agarose), with 200-bp (± 25 bp) fragments purified for downstream sequencing.
identifiers.namespace: SUB1313844
identifiers.id: Dietzia sp.111N12-1