ERC000011
ENA-CHECKLIST
2022-08-17T00:11:19Z
ENA-FIRST-PUBLIC
2022-08-17T00:11:19Z
ENA-LAST-UPDATE
SAMEA7192472
External Id
Nanjing Medical University
INSDC center name
2022-08-17T00:11:19Z
INSDC first public
2022-08-17T00:11:19Z
INSDC last update
public
INSDC status
ERS4952702
SRA accession
E-MTAB-9486:WT1
Submitter Id
10
day
age
ArrayExpress
broker name
house mouse
common name
Protocols: Euthanize mice using carbon dioxide (CO2) for 1-2 min or until breathing stops. Next, perform cervical dislocation on each mouse.Extract the mouse testis tissue and remove the capsule Total RNA was isolated using RNeasy mini kit (Qiagen, Germany). Strand-specific libraries were prepared using the TruSeq® Stranded Total RNA Sample Preparation kit (Illumina, USA)following the manufacturer’s instructions. Briefly, ribosomal RNA was removed from total RNA using Ribo-Zero rRNA removal beads. Following purification, the mRNA is fragmented into small pieces using divalent cations under 94℃ for 8 min. The cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers. This is followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragments then go through an end repair process, the addition of a single ‘A’ base, and then ligation of the adapters. The products are then purified and enriched with PCR to create the final cDNA library. Purified libraries were quantified by Qubit® 2.0 Fluorometer (Life Technologies, USA) and validated byAgilent 2100 bioanalyzer (Agilent Technologies, USA) to confirm the insert size and calculate the mole concentration.
description
juvenile stage
developmental stage
wild type genotype
genotype
Mus musculus
organism
Mus musculus
organism
testis
organism part
Mus musculus
scientific_name
male
sex
C57BL/6J
strain
WT1
title