Total RNA (10 μg/sample) was isolated and used to synthesize cDNA as described previously (13 (link)). The xfeA transcript in cDNA samples was sequenced using the c T408 F/R primers (Table S2). The sequencing chromatograms were analyzed with Chromas Lite (Technelysium, Brisbane, Australia), and the frequency of editing was estimated by ratiometric A/G measurement as described (13 (link)).
The EasyPure RNA kit was used to purify RNA as recommended (TransGen Biotech), and 1 μg of RNA was used to synthesize cDNA with the Magic 1st cDNA synthesis kit (Magic Biotech, Hangzhou, China). The cDNA product (20 μl) was diluted to 100 μl and used for qRT‐PCR with Magic SYBR green qPCR mix (Magic Biotech) and the ABI 7500 quantitative PCR system (Applied Biosystems, Foster City, CA). Expression was normalized with rpoD using the ΔΔCT method as described (13 (link)). Experiments included three independent biological replicates.
The EasyPure RNA kit was used to purify RNA as recommended (TransGen Biotech), and 1 μg of RNA was used to synthesize cDNA with the Magic 1st cDNA synthesis kit (Magic Biotech, Hangzhou, China). The cDNA product (20 μl) was diluted to 100 μl and used for qRT‐PCR with Magic SYBR green qPCR mix (Magic Biotech) and the ABI 7500 quantitative PCR system (Applied Biosystems, Foster City, CA). Expression was normalized with rpoD using the ΔΔCT method as described (13 (link)). Experiments included three independent biological replicates.
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