Get tips on using LC3B Antibody Kit for Autophagy to perform Autophagy assay cell type - Goblet cells
Get tips on using LC3B Antibody Kit for Autophagy to perform Autophagy assay cell type - SAE cells
Get tips on using LC3B Antibody Kit for Autophagy to perform Autophagy assay cell type - K562 cells
Get tips on using Guava Nexin Annexin V Assay to perform Apoptosis assay cell type - HeLa cells
Get tips on using Gibco™IMDM, powder to perform Stem cell Differentiation media hPSCs or iPSCs differentiation into Lung progenitor cells
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 Gibco™DMEM/F-12 to perform Stem cell culture media Human intestinal stem cells/organoids
Get tips on using OneDay ChIP kit to perform ChIP Human - Kupffer Cells
Get tips on using Anti-LC3B antibody produced in rabbit to perform Autophagy assay cell type - Paneth cells
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