|
| Status |
Public on Sep 01, 2016 |
| Title |
wt rspd rep1 |
| Sample type |
SRA |
| |
|
| Source name |
WT_round spermatids
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
age: post natal day 21
genotype: wild type
tissue: testes
tissue: round spermatids
|
| Treatment protocol |
6-8 week-old WT and Miwi KO female mice were each injected with 5IU of pregnant mare’s serum gonadotropin (PMSG), followed by injection of 5IU of human chorionic gonadotropin (hCG) 48h apart. Mature oocytes were collected from oviducts 14–16h after hCG injection and cumulus cells were removed by treatment with 0.1% bovine testicular hyaluronidase in HEPES-CZB at 37°C for 2–3 min. Cumulus-free oocytes were rinsed and kept in CZB at 37°C under 5% CO2 in air before ROSI.
|
| Growth protocol |
All animal work was performed following the protocol approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Nevada, Reno. Mice were housed and maintained under specific pathogen-free conditions with a temperature- and humidity-controlled animal facility in the University of Nevada, Reno. Global Miwi KO mouse line was recovered using cryopreserved sperm, purchased from the Mutant Mouse Resource & Research Center (MMRRC) (Item# 029995-MU-SPERM) in the Genetic Engineering Center of the University of Nevada, Reno. All mice used in this study were on the C57BL/6J background.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Small RNA was isolated from round spermatids using the mirVana RNA isolation kit (Ambion) according to the manufacturer’s instructions. RNA quality and quantity were assessed using the Agilent 2100 Bioanalyzer.
Libraries were prepared using the Ion Total RNA-Seq Kit v2 (Invitrogen)
|
| |
|
| Library strategy |
ncRNA-Seq |
| Library source |
transcriptomic |
| Library selection |
size fractionation |
| Instrument model |
Ion Torrent Proton |
| |
|
| Description |
wild type purified round spermatids
|
| Data processing |
Torrent Suite
Cutadapt remove adaptor sequence
The reads were mapped to reference small RNA file though our house pipeline
The mapped reads were counted though featureCount
The counts were normalize though Deseq2
Genome_build: mm9
Supplementary_files_format_and_content: counts txt file
|
| |
|
| Submission date |
Aug 31, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Wei Yan |
| E-mail(s) |
wyan@medicine.nevada.edu
|
| Phone |
(775) 784-7765
|
| Organization name |
University of Nevada
|
| Department |
Physiology
|
| Lab |
Wei Yan Lab
|
| Street address |
1664 N. Virginia Street
|
| City |
reno |
| State/province |
NV |
| ZIP/Postal code |
89557 |
| Country |
USA |
| |
|
| Platform ID |
GPL18635 |
| Series (1) |
| GSE86319 |
Paternal pachytene piRNAs are not required for fertilization, embryonic development and paternal epigenetic inheritance in mice |
|
| Relations |
| BioSample |
SAMN05721815 |
| SRA |
SRX2071184 |
