CRISPR Mouse Deletion RAW 264.7

Get tips on using Macrolide – R/MG ELITe MGB®Kit to perform Cell Culture Contamination Detection Kit Mycoplasma

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Get tips on using IMAGEN™ Respiratory Virus Screen Kit using Direct Immunofluorescence Assay to perform Cell Culture Contamination Detection Kit Virus

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Get tips on using PrepSEQ™ 1-2-3 Mycoplasma Nucleic Acid Extraction Kit to perform Cell Culture Contamination Detection Kit Mycoplasma

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Get tips on using IMAGEN™ Respiratory Syncytial Virus Kit (RSV) using Direct Immunofluorescence Assay 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.