2024-11-18
ENA first public
ERC000011
ENA-CHECKLIST
Jospe Carreras Leukemia Research Institute
INSDC center name
public
INSDC status
ERS19301987
SRA accession
E-MTAB-14048:NEKOd7_1
Submitter Id
ArrayExpress
broker name
ReNcell VM Human Neural Progenitor
cell_line
not collected
collection date
human
common name
Protocols: Samples were collected from cell cultures maintained under standard growing conditions for neural progenitor cells. Cells were pelleted and total RNA extraction was carried out with the Maxwell (Promega) technology.  Stranded mRNA Library construction procedure are listed below:  1) mRNA enrichment and purification:  Oligo dT Selection (mRNA enrichment): Use oligo dT beads to enrich mRNA with poly A tail.  2) RNA fragment and reverse transcription  Fragment the RNA and first-strand cDNA was generated using random N6-primed reverse transcription, followed by a second-strand cDNA synthesis with dUTP instead of dTTP.  3) End repair, add A and adaptor ligation  The synthesized cDNA was subjected to end-repair and then was 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments;  4) PCR amplification  Prior to PCR amplification, the dUTP-marked strand is selectively degraded by Uracil-DNA-Glycosylase(UDG). The remaining strand is amplified to generate a cDNA library suitable for sequencing. Many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer;  5) Single strand separation and cyclization  Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and DNA ligase;  6) DNA nanoball synthesis
description
Rett syndrome
disease
NEAT1 knockout
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
1
replicate
Homo sapiens
scientific_name
NEKOd7_1
title