Get tips on using Ambion™ RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE to perform RNA isolation / purification Tissue - Human Kidney
Get tips on using Beta-Lactamase Activity Assay Kit to perform Reporter gene assay β-lactamase substrates - Burkholderia cepacia complex
Get tips on using Ambion™ RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE to perform RNA isolation / purification Tissue - rat kidney tissue
Get tips on using Ambion™ RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE to perform RNA isolation / purification Tissue - human kidney tissue
Get tips on using Ambion™ RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE to perform DNA isolation / purification Bacteria - Gram positive Lactobacillus
Get tips on using Active rat PAI-1 functional assay ELISA kit to perform ELISA Rat - Serpin E1/PAI-1
Get tips on using Rat plasminogen activator inhibitor 1,PAI1 ELISA Kit to perform ELISA Rat - Serpin E1/PAI-1
Get tips on using Mitochondrial ROS Activity Assay Kit (Deep Red Fluorescence) to perform ROS assay cell type - mouse cardiomyocytes
Get tips on using ViralSEQ™ Quantitative Sf-Rhabdovirus Kit with PrepSEQ™ Nucleic Acid Sample Prep Kit to perform Cell Culture Contamination Detection Kit Virus
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.
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