siRNA / miRNA gene silencing Human HeLa EPAS-1

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 RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human umbilical vein smooth muscle cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human umbilical artery smooth muscle cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human renal proximal tubular epithelial cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human pulmonary artery smooth muscle cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human coronary artery smooth muscle cells

Get tips on using RNeasy Micro Kit to perform RNA isolation / purification Cells - primary human hair follicle dermal papilla cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human hair follicle dermal papilla cells

Get tips on using Minimum Essential Medium Eagle to perform Stem cell culture media Human bone mesenchymal stem cell (BMSC)

Get tips on using Atg5 (D5F5U) Rabbit mAb to perform Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)