KO 2
Mus musculus
Protocols: Slc7a5 fl/fl mice harbouring two copies of the Slc7a5 targeted allele (exon 1 of Slc7a5 flanked with two loxP sites), were crossed with a mouse line ubiquitously expressing cre recombinase under the Bal1 promoter (Bal1-cre) to obtain a global Slc7a5 knockout mouse line (Poncet et al, 2014). Heterozygous Slc7a5+/- C57Bl/6 mice were viable and fertile. However, Slc7a5+/- inter-crosses did not produce any Slc7a5-null live mice. Here, heterozygous Slc7a5+/- mice were crossed to generate wild-type and Slc7a5-null embryos isolated at stage E8.5. Mouse colonies were breed, mice sacrificed and embryos isolated following Home Office guidelines (PPL 60/04454 or PPL 60/3455 and/or 60/4118). E8.5 embryos with 8 to 10 somites were dissected in cold PBS and lysed individually with Trizol reagent. Embryos were stage matched with respect to somite number and only male embryos were used to avoid potential sex-related variations in transcriptome. Allantois was kept for genotyping and sexing the embryos by PCR (McFarlane et al, 2013). Total RNA was extracted using the RNeasy mini kit (Qiagen). Samples were DNase treated 'on-column' using RNase-free DNase (Qiagen). Quality and quantity of total RNA were measured using a Qubit fluorometer (Invitrogen). Following quality control, 5 wild type and 5 Slc7a5-null male embryos were selected as biological replicates. Samples started with 300ng total RNA per embryo to which spike-ins (ERCC, 2ul of 1:100 dilution) were added. We used a TruSeq RNA v2 kit (Illumina RS-122-2001) to prepare the cDNA library following manufacturer's instructions; polyA RNA selection was performed and fragment and random priming followed by cDNA synthesis, ligation and x10 rounds of PCR amplification.
ENA first public
2018-06-01
ENA last update
2018-01-09
External Id
SAMEA104458461
INSDC center alias
Division of Computational Biology, University of Dundee
INSDC center name
Division of Computational Biology, University of Dundee
INSDC first public
2018-06-01T17:03:03Z
INSDC last update
2018-01-09T13:38:32Z
INSDC status
public
Submitter Id
E-MTAB-6336:KO 2
broker name
ArrayExpress
common name
house mouse
developmental stage
embryonic day 8.25
genotype
Bal1-cre / Slc7a5 flox/flox
organism part
whole organism
phenotype
Slc7a5 knockout
sample name
E-MTAB-6336:KO 2
sex
male
strain
C57BL/6