siRNA / miRNA gene silencing Human IMR-90

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines Neuro2a

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines L02

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines H9C2

Get tips on using jetPEI® DNA transfection, HTS application to perform DNA transfection Mammalian cells - Immortalized cell lines Chang Liver cells

Get tips on using jetPEI® DNA transfection, HTS application to perform DNA transfection Mammalian cells - Immortalized cell lines MDA-MB-231

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines Huh7

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines HepG2

Get tips on using Recombinant Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063) to perform Immunohistochemistry Mouse - ERα

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines PANC-1

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.