Experiment information
Accession CRX020694
Organism Ipomoea batatas
Title C11_T39
BioProject PRJCA000642
BioSample SAMC017536
Platform Illumina HiSeq 2500
Library name Construction protocol Strategy Source Selection Layout
C11_T39 A total amount of 3 ?g RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext®Ultra™ RNA Library Prep Kit for Illumina®(NEB, USA) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer?5X?. First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase?RNase H-?. Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H . Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3’ ends of DNA fragments, NEBNext Adaptor with hairpin loop structure were ligated to prepare for hybridization. In order to select cDNA fragments of preferentially 150~200 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA). Then 3 ?l USER Enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37°C for 15 min followed by 5 min at 95°C before PCR. Then PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system. RNA-Seq TRANSCRIPTOMIC cDNA PAIRED
Processing Planned read length (bp) for mate 1: 100
Planned read length (bp) for mate 2: 100
Insert size (bp): 300
Release date2017-11-22
Run Accession Release date Run Data File Information
File nameFile size (MB)
CRR022913 2017-11-22 Batata_C11--T39-I_good_1.fq.gz
SubmitterYonghai Luo (yonghailuo@foxmail.com)
OrganizationsJiangsu Normal University
Date submitted2017-11-22