title: GSM1849333: Hind wing bud RNA sequence left and right reads; Bombyx mori; RNA-Seq
library_construction_protocol: Fore and Hind wing buds were isolated from second and third thoracic segments of the fourth instar larvae after temporarily aneasthetizing them on ice. The Wing buds were plunged directly into liquid Nitrogen and stored at -80C prior to trizol extraction of the RNA. The extracted RNA was processed through Agen court purification before proceedng for library preperation. Transcriptome library for sequencing was constructed according to the Illumina TruSeq RNA library protocol outlined in “TruSeq RNA Sample Preparation Guide” (Part # 15008136; Rev. A; Nov 2010). Briefly, the Total RNA was fragmented for 4 minutes at elevated temperature (94oC) in the presence of divalent cations and reverse transcribed with Superscript II Reverse transcriptase by priming with Random Hexamers. Second strand cDNA was synthesized in the presence of DNA Polymerase I and RnaseH. The cDNA was cleaned up using Agencourt Ampure XP SPRI beads (Beckman Coulter). Illumina Adapters were ligated to the cDNA molecules after end repair and addition of “A”- base. SPRI cleanup was performed after ligation. The library was amplified using 11 cycles of PCR for enrichment of adapter ligated fragments. The prepared library was quantified using Nanodrop and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent)
attributes.tag: GEO Accession
attributes.value: GSM1849333