rna-isolation-purification-cells-immortalized-ovcar-3

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Tissue - lung

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Tissue - brain

Get tips on using DNeasy PowerLyzer PowerSoil Kit (100) to perform DNA isolation / purification Insects

Get tips on using MicroRNA isolation kit to perform CRISPR

Get tips on using MicroRNA isolation kit to perform siRNA / miRNA gene silencing Rat - IEC-6 HuR

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 Quant-iT™ RiboGreen™ RNA Assay Kit to perform RNA quantification Fuorimetric - human metastatic melanoma cells