title: 11T
organism: Homo sapiens
description: Protocols: Prostate tumor and adjacent normal tissues from 14 patients obtained from Shanghai Changhai Hospital were selected as the discovery cohort for RNA sequencing. RNA-seq libraries were constructed using the Illumina kit following the manufacturer's standard protocol. The poly(A) containing RNA were enriched from 5ug total RNA and fragmented to 200-250nt in size via fragmentation buffer at 70 C for 90 seconds. The first cDNA strand was synthesized with random hexamer primers using Super transcriptase II and the second-strand synthesis was performed by incubation with RNase H and DNA polymerase. End-repair was performed using Klenow polymerase, T4 DNA polymerase and T4 polynucleotide kinase. The end-repaired cDNA fragments were 3- adenylated with Klenow exo- and dATP, followed by Illumina PE adapters ligation on both ends. After recovery from agarose gel, the ligation products were amplified by 15 cycles of PCR with Phusion polymerase. The final size and yield of each library was validated on Agilent Technologies 2100 Bioanalyzer and q-PCR separately, and was diluted to 2.5pM for sequencing on a single lane of an Illumina HiSeq2000 flowcell.
attributes.tag: External Id
attributes.value: SAMEA1027588
attributes.tag: INSDC center alias
attributes.value: BGI
attributes.tag: INSDC center name
attributes.value: Beijing Genome Institute
attributes.tag: INSDC first public
attributes.value: 2011-12-31T17:00:24Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T15:25:30Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-567:11T
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: human
attributes.tag: sample name
attributes.value: E-MTAB-567:11T