GSM2410209: adult rep2; Nesidiocoris tenuis; RNA-Seq
RNA was isolated from adults using the TRIzol (Invitrogen) reagent by following the company manual. Poly(A) mRNA was isolated from 20 µl total RNA using Oligo (dT) magnetic beads and then was broken into short fragments (about 200bp) in the presence of fragmentation buffer at 94 °C for 5 min. The short fragments (about 200 bp) were used as templates for first-strand cDNA synthesis using random hexamer-primers. Subsequently, second-strand cDNAs were synthesized using buffer, dNTPs, RNaseH, and DNA polymerase I. After purification of short fragments, samples were then washed with EB buffer for end reparation and single nucleotide adenine addition. Finally, the short fragments were connected to sequencing adapters. Suitable fragments (about 200bp), as judged by agarose gel electrophoresis, were enriched with PCR amplification to prepare the sequencing library.
GEO Accession
GSM2410209