RNA was isolated from homogenates of freshly isolated rat PAs, mesenteric arteries, aorta, brain, and cultured PASMCs. Reverse transcription of the RNA was carried out as previously described (16 (link)). RT-PCR primer pairs for rat sulfide quinone reductase-like (SQRDL) were designed using Primer3web version 4.0 (http://primer3.ut.ee ) and synthesized by Sigma-Aldrich (St. Louis, MO). SQRDL (Accession No. BC158559 ) primers were sense CTGCAGGACTTCAAGGAAGG and antisense CTCTCCCGAATGATCTCCTG. PCR was carried out using PuReTaq Ready-To-Go PCR Beads (GE Healthcare, Piscataway, NJ), and the PCR products (reaction equivalent on 20 ng reverse transcribed RNA) were analyzed by electrophoresis on 2.8% agarose gels run in TAE buffer (National Diagnostics, Hessle, UK) with PhiX174 DNA/HinfI Marker (Thermoscientific, Waltham, MA).
Tae buffer
Manufactured by National Diagnostics
Sourced in United Kingdom
TAE buffer is a common buffer solution used in molecular biology and biochemistry laboratories. It is primarily used as a buffer for agarose gel electrophoresis, a technique employed to separate and analyze DNA, RNA, or protein samples. TAE buffer maintains a specific pH range to ensure the appropriate migration and separation of these biomolecules during electrophoresis.
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Lab products found in correlation
2 protocols using tae buffer
1
Quantifying Sulfide Quinone Reductase Expression
2
Reverse Transcription and RT-PCR Analysis of Rat IPA
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RNA was isolated from homogenates of freshly isolated rat IPAs and reverse transcription carried out as previously described (Knock et al. 2008 (link)). RT-PCR primer pairs were designed using Primer3web version 4.0 (http://primer3.ut.ee ) and synthesized by Sigma-Aldrich (St Louis, MO, USA). MPT (accession no. NM_138843): sense CCTTCATCAAGACCCACGAG, antisense TGGACAGGTCCACCTTCTTC; CβS (accession no. NM_012522): sense ATGCTGCAGAAAGGCTTCAT, antisense CAACACCAAACACCATCAG; CγL (accession no. NM_017074): sense GGTTTTGTATACAGCCGCTCTGG, antisense TGCAAATGACTTCATCTCCTGCT. PCR was carried out using PuReTaq Ready-To-Go PCR Beads (GE Healthcare, Piscataway, NJ, USA) and the PCR products (reaction equivalent on 20 ng reverse transcribed RNA) were analysed by electrophoresis on 2.8% agarose gels run in TAE buffer (National Diagnostics, Hessle, UK) with PhiX174 DNA/HinfI Marker (Thermoscientific Inc., Waltham, MA, USA).
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