2025-12-13
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA115080413
External Id
NewStem LTD
INSDC center name
2024-01-02T17:01:31Z
INSDC last update
public
INSDC status
ERS17694156
SRA accession
E-MTAB-13685:Sample 3
Submitter Id
ArrayExpress
broker name
haploid embryonic stem cells
cell_type
not collected
collection date
human
common name
Protocols: Wash cells with PBS x2. Trypsinize (TrypLE Select) the wells with the regular protocol. Spin down the cells after neutralizing the trypsin with a serum-containing medium. Count cellsfreeze pellet gSYNC DNA Extraction Kit (GS100) prepare mix inside a PCR hood, after cleaning the surface with DNase Away and70% EtOH. If possible, we recommend setting up at least 4 parallel PCRs for a given sample.Final contents of each reaction:• 10 μL 10x reaction buffer• 8 μL dNTP• 0.5 μL P5 primer mix, 100 μM• 1.5 μL ExTaq polymerase• up to 10 μg of genomic DNA or 200 ng of plasmid DNA• 10 μL of P7 primer 5 μM• up to 100 μL with water1. Make a master mix of reaction buffer, dNTP, P5 primer mix, taq polymerase, and water.Aliquot into a PCR plate.2. Add template DNA.3. Add a unique P7 primer to barcode each individual reaction.Thermal cycler parametersWait for block to reach 95°C before adding samples.1. 95°C, 1 minute2. 95°C 30 seconds (denaturation)3. 53°C 30 seconds (annealing)4. 72°C 30 seconds (extension)Back to step 2, total of 28 cycles5. 72°C 10 minutes6. 4°C forever
description
embryo
developmental stage
TP53 KO
genetic modification
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
inner cell mass
organism part
Homo sapiens
scientific_name
14
day
time
Sample 3
title