description: Protocols: Human embryonic kidney 293 cells were cultured in RPMI containing 10% foetal bovine serum and penicillin/streptomycin at 37°C, 5% CO2. The cells were transfected with the indicated plasmids or siRNAs using JetPRIME (Polyplus) according to manufacturer's recommendations. Total RNA was prepared using TRIzol (Life Technologies) and a standard protocol. RNA was resuspended in RNase-free water and quantified with NanoDrop (ThermoScientific). RNA samples were treated with DNaseI (Promega) and reverse transcribed with a gene-specific primer using Maxima H Minus Reverse Transcriptase (Thermo Scientific). Complementary DNA was amplified in nested PCR reactions using taxon-specific PCR primers shown in Table S1 and Figure 3B. PCR products were separate on agarose gels to check their expected size. PCR products were extracted from agarose gel slices using GeneJET Gel Extraction Kit (ThermoScientific) and quantified by NanoDrop. cDNA samples were diluted and the integrity and concentration of the DNA was confirmed by Agilent 2100 Bioanalyzer (Agilent Technologies). Ampli-seq libraries were prepared using proprietary adapter ligation and amplification (Eurofins Genomics).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2021-02-07T04:10:42Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-08-10T15:36:30Z
attributes.tag: External Id
attributes.value: SAMEA7171667
attributes.tag: INSDC center name
attributes.value: University of Southampton School of Medicine
attributes.tag: INSDC first public
attributes.value: 2021-02-07T04:10:42Z
attributes.tag: INSDC last update
attributes.value: 2020-08-10T15:36:30Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9412:Sample 7
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: HEK293T
attributes.tag: common name
attributes.value: chicken
attributes.tag: genotype
attributes.value: expression of chicken KGDH intron lariat junctions
attributes.tag: sample name
attributes.value: E-MTAB-9412:Sample 7
attributes.tag: scientific_name
attributes.value: Gallus gallus