shRNA gene silencing Human HEK 293T

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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

INTERFERin® Product

Get tips on using INTERFERin® to perform siRNA / RNAi /miRNA transfection Mouse - B16 Polymer / lipid

Products Polyplus transfections INTERFERin®

HiPerFect Transfection Reagent Product

Get tips on using HiPerFect Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - AR42J Lipid based