siRNA / miRNA gene silencing Rat MTLn3 (rat mammary adenocarcinoma breast cancer cell line)

Get tips on using Minimum Essential Media (MEM) to perform Mammalian cell culture media MDCK

Get tips on using Minimum Essential Media (MEM) to perform Mammalian cell culture media Vero

Get tips on using Minimum Essential Medium Eagle to perform Mammalian cell culture media Vero

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 ATP Assay to perform Cell cytotoxicity / Proliferation assay cell type - adipose stem cells

Get tips on using Human Synoviocyte Growth Medium to perform Mammalian cell culture media HFLS-OA

Get tips on using Minimum Essential Media (MEM) to perform Mammalian cell culture media BHK-21

Get tips on using LC3B Antibody #2775 to perform Autophagy assay cell type - Mast cell

Get tips on using SPLInsert™ Hanging to perform Cell migration / Invasion cell type - HaCat