cdna-synthesis-tissue

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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 Tissue

Get tips on using Tetro cDNA Synthesis Kit to perform cDNA synthesis Tissue

Products Bioline Tetro cDNA Synthesis Kit

Get tips on using qScript cDNA Synthesis Kit to perform cDNA synthesis Tissue

Products Quantabio qScript cDNA Synthesis Kit

Get tips on using iScript cDNA Synthesis Kit to perform cDNA synthesis Tissue

Products Bio-Rad Laboratories iScript cDNA Synthesis Kit

Get tips on using First-Strand cDNA Synthesis Kit to perform cDNA synthesis Tissue

Products GE Healthcare Life Sciences First-Strand cDNA Synthesis Kit

Get tips on using ReadyScript® cDNA Synthesis Mix to perform cDNA synthesis Tissue

Products Sigma-Aldrich ReadyScript® cDNA Synthesis Mix

Get tips on using Transcriptor High Fidelity cDNA Synthesis Kit to perform cDNA synthesis Tissue

Products Sigma-Aldrich Transcriptor High Fidelity cDNA Synthesis Kit

Get tips on using PrimeScript™ 1st strand cDNA Synthesis Kit to perform cDNA synthesis Tissue

Products Takara Bio Inc PrimeScript™ 1st strand cDNA Synthesis Kit

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

Get tips on using iScript cDNA Synthesis Kit to perform cDNA synthesis Yeast

Products Bio-Rad Laboratories iScript cDNA Synthesis Kit

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