2024-11-22
ENA first public
ERC000011
ENA-CHECKLIST
EMBL
INSDC center name
public
INSDC status
ERS22445200
SRA accession
E-MTAB-14636:SpermRNAyFVBCTL033
Submitter Id
10 to 11
week
age
ArrayExpress
broker name
not collected
collection date
house mouse
common name
Protocols: Male mice were randomly assigned to one of three treatment groups: Control (CON), non-absorbable antibiotics (nABX), or low protein high sugar diet (LPHS). Young (3 weeks) and mature (12-13 months) mice in the CON group received water and standard rodent diet, nABX mice received antibiotics in their water (Neomycin, Bacitracin, Pimaricin) along with the standard diet, and LPHS mice were given a low-protein, high-sugar (LPHS) diet and water. Cauda epididymides from FVB and C57BL/6J control and treated mice were dissected, cleaned, and incubated in M2 media to allow spermatozoa to swim out. After 30 minutes at 37C and 5% CO2, remaining sperm was released by gently pinching tissues, followed by an additional 10-minute incubation. The sperm suspension was filtered, centrifuged, and resuspended in somatic cell lysis buffer on ice for 20 minutes to remove any somatic cell contamination. After further washes and centrifugation, sperm pellets were dried and stored at -80C. Sperm RNA was extracted using an optimized mirVana miRNA isolation kit. Briefly, spermatozoa were lysed in mirVana lysis buffer with a tissue homogenizer. The lysed solution underwent organic extraction and RNA isolation following the mirVana protocol, and including an Acid-Phenol: Chloroform step. RNA was precipitated, purified, washed, and eluted with nuclease-free water, and its concentrations were measured using the TapeStation, ranging from 1000 to 8000 pg/µL. The RNA was stored at -80C. Libraries were constructed using the NextFlex Small RNA-Seq Kit v4 by PerkinElmer, with varying concentrations of starting material but consistent volumes across samples.
description
not collected
geographic location (country and/or sea)
birth
initial time point
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
sperm
organism part
Mus musculus
scientific_name
FVB
strain
SpermRNAyFVBCTL033
title
CON
treatment