siRNA / miRNA gene silencing Human IMR-90

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.

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.

Get tips on using Anti Type X Collagen (raised against rat) pAb (Rabbit, Antiserum) to perform Immunohistochemistry Mouse - Col X

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines MDA-MB-231

Get tips on using MasterPure™ Complete DNA and RNA Purification Kit to perform DNA isolation / purification Cells - Immortalized cell lines C2C12

Get tips on using Wilms' Tumor 1 (WT1) Protein, Clone 6F-H2 to perform Immunohistochemistry Wilms Tumor 1 (WT1) - Rabbit Mouse -NA-

Get tips on using Purified Mouse Anti-SV40 Large T Antigen Clone PAb 101 (RUO) to perform Immunohistochemistry Mouse - SV40

Get tips on using FitAmp Blood and Cultured Cell DNA Extraction Kit to perform DNA isolation / purification Cells - Immortalized cell lines SH-SY5Y

Get tips on using anti-alpha-Smooth Muscle Actin mouse monoclonal, ASM-1 to perform Immunohistochemistry Alpha smooth muscle Actin - Mouse -NA- -NA-

Get tips on using Purified anti-Tubulin β-3 (TUBB3) Antibody (Previously Covance catalog# PRB-435P) to perform Immunohistochemistry Mouse - TUBB3