Digoxin high prime dna labeling and detection starter kit 1

The genomic DNAs were isolated using the cetyl trimethyl ammonium bromide method [41] (link). The restriction digestion, gel electrophoresis, ligation reactions, and PCR were all carried out using standard procedures. Southern blotting was performed using the digoxin high-prime DNA labeling and detection starter kit I (Roche) following the manufacturer's instructions.
Total RNA was isolated using the Trizol reagent (Invitrogen, Carlsbad, CA, USA), and used as a template to synthesize cDNA using AMV Reverse Transcriptase (Takara Bio, Otsu, Japan). The abundance of MoPEX19 transcripts was analyzed using primer pair 19RTF/19RTR on the 7500 Fast Real-Time System (Applied Biosystems, Foster, CA, USA) by calculating the average threshold cycle (Ct) value, which was normalized to that of Tubulin gene (MGG_00604). The relative abundance of gene expression at deferent developmental stages was compared with that in fresh conidia. Three replicates were performed for each sample.

The genomic DNA was isolated using the cetyl trimethyl ammonium bromide method [48 (link)]. Total RNA was prepared using the Trizol reagent (Invitrogen, Carlsbad, CA, USA), and used to synthesize the cDNA using AMV Reverse Transcriptase (Takara Bio, Otsu, Japan). PCR, Restriction digestion, gel electrophoresis and ligation reactions were performed using standard procedures. Transcript abundance was analyzed by quantitative PCR on the 7500 Fast Real-Time System (Applied Biosystems, Foster, CA, USA) with the β-tubulin gene (MGG00604) for normalization. Southern blotting was performed using the digoxin high-prime DNA labeling and detection starter kit I (Roche).