Experiment information
Accession CRX044393
Organism Homo sapiens
Title TMEM30A-KO1
BioProject PRJCA001319
BioSample SAMC063942
Platform Illumina HiSeq 2500
Library name Construction protocol Strategy Source Selection Layout
After extraction of total RNA from HRECs using RNeasy Mini kits (QIAGEN), the RNA purity was checked using a K5500 spectrophotometer (Kaiao, Beijing,China). RNA integrityand concentration were assessed using an RNA Nano 6000 Assay Kit of a Bioanalyzer 2100 system (Agilent Technologies,CA).Forlibrarypreparation,atotalof2 μgRNApersamplewas used as input material for the RNA sample preparations. Sequencing libraries were generated using NEB Next, Ultra RNA Library Prep Kit for Illumina® (Cat# E7530L, NEB, USA), following the 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 conditions in NEBNext First Strand Synthesis Reaction Buffer (5×). First-strand cDNA was synthesized using random hexamer primers and RNase H. Second-strand cDNA synthesis was subsequently performed using dNTPs,DNApolymeraseI andRNaseH.Thelibraryfragmentswerepurified with QiaQuick PCR kits and elution with EB buffer, and then terminal repair, A-tailing and addition of an adapter were implemented. The RNA concentration of the library was measured using a Qubit RNA Assay Kit in Qubit 3.0 to preliminarily quantify and then dilute to 1 ng/μl. Insert size was assessed using the Agilent Bioanalyzer 2100 system (Agilent Technologies), and the qualified insert size was accurately quantified using a StepOnePlus™ Real-TimePCRSystem(libraryvalidconcentration>10 nM).Theclusteringof the index-coded samples was performed on a cBot cluster generation system using a HiSeq PE Cluster Kit v4-cBot-HS (Illumina, CA, USA) according to the manufacturer’s instructions. After cluster generation, the libraries were sequenced on an Illumina HiSeq 2500 platform, and 150 bp paired-end reads were generated. All gene expression values from RNA-seq were changed to a log2valueandanalyzedfurther. RNA-Seq TRANSCRIPTOMIC unspecified PAIRED
Processing Planned read length (bp) for mate 1: 150
Planned read length (bp) for mate 2: 150
Insert size (bp): 300
Planned read length (bp): 150
Release date2019-03-14
Run Accession Release date Run Data File Information
File nameFile size (MB)
CRR049205 2019-03-14 KO1_R1.fq.gz
Submittershan zhang (zhangshanshan@std.uestc.edu.cn)
OrganizationsSichuan Provincial People
Date submitted2019-03-11
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