Experiment information
Accession CRX101341
Organism Homo sapiens
Title RNA-seq of PBMC: healthy donor 2
BioProject PRJCA002326
BioSample SAMC149135
Platform Illumina NovaSeq 5000
Library
Library name Construction protocol Strategy Source Selection Layout
One microgram of total RNAs extracted from PBMCs (Ficoll preparation) were used as input. Messenger RNAs were purified using oligo-dTs covalently coupled magnetic beads. Then, the RNAs were fragmented into small pieces by heating. First strand of cDNA was synthesized in the presence of specific chemicals to ensure that only RNAs were used as template. Double strand cDNAs were purified with Agencourt AMPure XP beads after the reaction. DNA library was constructed through end-repair, adaptor-ligation and PCR amplification. The intermediate products were size-selected after the adaptor-ligation using two rounds of Agencourt AMPure XP beads. Qualified double strand DNA library was transformed into single-stranded circular DNA library through DNA-denaturation and circularization. DNA nanoballs (DNBs) were generated from single-stranded circular DNA using rolling circle amplification (RCA). The DNBs were qualified using Qubit 2.0. Qualified DNBs were loaded on the flow cell and sequenced. RNA-Seq TRANSCRIPTOMIC PolyA PAIRED
Processing Planned read length (bp) for mate 1: 150
Planned read length (bp) for mate 2: 150
Insert size (bp): 145
Release date2020-04-01
Run
Run accession Run data file information
File nameFile size (MB)
CRR125445 CRR125445_f1.fastq.gz
CRR125445_r2.fastq.gz
3,682.52
3,836.07
SubmitterYu Zhou (yu.zhou@whu.edu.cn)
OrganizationWuhan University
Date submitted2020-03-18