title: PolyA+_testis
organism: Mus musculus
description: Protocols: An 8-week-old C57/BL6 mouse purchased from Charles River was sacrificed, from which brain, heart, kidney, liver, lung, muscle (biceps femoris), spleen and testis were dissected. The tissues were homogenised in Trizol and total RNA was extracted. Any remaining DNA was removed with the Qiagen RNase-free DNase kit (ref. 79254) and total RNA was cleaned up with a Qiagen RNeasy mini kit (ref. 74104) column. Then, poly-A+ RNA was extracted from the total RNA using the Qiagen Oligotex mRNA mini kit (ref. 70022). Reverse transcription was performed with the Invitrogen SuperScript III first-strand synthesis kit (ref. 18080-051) using random hexamers for priming. PCR amplifications were performed using 0.2 ng of initial poly A+ RNA (as measured before doing the reverse transcription) in a final volume of 12.5 uL with JumpStart REDTaq ReadyMix (Sigma-Aldrich) and a primer concentration of 0.4 uM. Reactions took place in 384-well plates format on an automatized Evoware platform (TECAN) combined with a Tetrad2 thermocycler (Bio-Rad) that allows processing four plates in parallel. Because monoexonic amplification is sensitive to genomic DNA contaminations, monoexonic models were assessed by amplification of cDNA in which a dNTP analog was incorporated using the mRNA Selective PCR Kit (TAKARA).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2015-01-22T17:03:08Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T22:45:00Z
attributes.tag: External Id
attributes.value: SAMEA3216849
attributes.tag: INSDC center name
attributes.value: Wellcome Trust Sanger Institute
attributes.tag: INSDC first public
attributes.value: 2015-01-22T17:03:08Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T22:45:00Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-3261:PolyA+_testis
attributes.tag: age
attributes.value: 8 weeks
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: organism part
attributes.value: testis
attributes.tag: sample name
attributes.value: E-MTAB-3261:PolyA+_testis
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57BL/6