2024-06-15
ENA first public
ERC000011
ENA-CHECKLIST
University of Southern California Leonard Davis School of Gerontology
INSDC center name
public
INSDC status
ERS20166423
SRA accession
E-MTAB-14132:unc-25p::xbp-1s_1
Submitter Id
1
day
age
ArrayExpress
broker name
not collected
collection date
Protocols: For RNA isolation, all RNa collection was performed at day 1 of adulthood. ~1000 animals were harvested from RNAi plates using M9. Animals were pelleted by gravity by allowing adult worms to settle to the bottom of the tube and aspirating off eggs and L1. Animals were washed and gravity settled 3x to remove a majority of progeny, then animals were placed into Trizol solution and worms were freeze/thawed 3x with liquid nitrogen with a 30 sec vortexing step between each freeze cycle. Standard bleaching protocol was used to synchronize animal populations. Animals were L1 arrested for a maximum of 16 hours to further synchronize populations. Animals were grown on HT115 bacteria from L1 until Day 1 of adulthood at 22°C (the restrictive temperature for our glp-4(bn2) worms and material was collected on Day 1. After the final thaw, chloroform was added at a 1:5 chlorform/trizol ratio and aqueous separation of RNA was performed via centrifugation in a heavy gel phase-lock tube (VWR, 10847-802). The aqueous phase was mixed 1:1 with isopropanol then applied to a Qiagen RNeasy Mini Kit (74106) and RNA purification was performed as per manufacturer’s directions.  Library construction was performed by Novogene using their standard pipeline.
description
adult
developmental stage
glp-4(bn2); GABAergic xbp-1s
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Caenorhabditis elegans
organism
Caenorhabditis elegans
organism
whole organism
organism part
Caenorhabditis elegans
scientific_name
hermaphrodite
sex
N2
strain
unc-25p::xbp-1s_1
title