RNA isolation / purification Yeast

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cDNA synthesis Yeast Experiment

The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.

RNA cDNA synthesis Yeast

Wizard® Genomic DNA Purification Kit Product

Get tips on using Wizard® Genomic DNA Purification Kit to perform DNA isolation / purification Yeast - Candida parapsilosis

Products Promega Wizard® Genomic DNA Purification Kit

Wizard® Genomic DNA Purification Kit Product

Get tips on using Wizard® Genomic DNA Purification Kit to perform DNA isolation / purification Yeast - Pichia pastoris

Products Promega Wizard® Genomic DNA Purification Kit

Total RNA Purification Kit Product

Get tips on using Total RNA Purification Kit to perform RNA isolation / purification Tissue - Mouse Liver