Western blot Secondary Antibody Goat Rabbit

Get tips on using Anti-Collagen, Type VII to perform Immunohistochemistry Collagen Type VII - Rabbit Human -NA-

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary rabbit aortic smooth muscle cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary rabbit aortic endothelial cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary rabbit skeletal muscle cells

Get tips on using APO-BRDU™ Kit (RUO) to perform TUNEL assay cell type - Rabbit synovial fibroblasts

Get tips on using Ki-67 Antigen, Clone MIB-1 to perform Immunohistochemistry Ki67 - Rabbit Mouse / Human -NA-

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary rabbit skeletal muscle-derived stem cells

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Rabbit eye retina/choroids

Get tips on using T-PER™ Tissue Protein Extraction Reagent to perform Protein isolation Tissue - Rabbit eye retina/choroids

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.