rna-isolation-purification-cells-immortalized-cos-7

Get tips on using Quant-iT™ RiboGreen™ RNA Assay Kit to perform RNA quantification Fuorimetric - mouse liver tissue

Get tips on using RediPlate™ 96 RiboGreen™ RNA Quantitation Kit to perform RNA quantification Fuorimetric - mouse adipose tissue

Get tips on using RediPlate™ 96 RiboGreen™ RNA Quantitation Kit to perform RNA quantification Fuorimetric - human brain tissue

Get tips on using Cell Lysis Buffer (10X) to perform Protein isolation Mammalian cells - THP-1

Get tips on using Cell Lysis Buffer (10X) to perform Protein isolation Mammalian cells - KC02-44D

Get tips on using NP40 Cell Lysis Buffer to perform Protein isolation Mammalian cells - BHK-21

Get tips on using RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - Rat_Mesenteric fat

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 RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - HepG2

Get tips on using RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - HEK293T