Protein Expression Prokaryotic cells A. tumefaciens

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - mouse mesenchymal stem cells

Get tips on using Mammary Epithelial Cell Growth Medium to perform 3D Cell Culture Media Human primary breast ephitelial cells-organoids

Get tips on using Mammary Epithelial Cell Growth Medium to perform 3D Cell Culture Media Human primary breast ephitelial cells-Mammospheres

Get tips on using Mesenchymal Stem Cell Chondrogenic Differentiation Medium to perform Stem cell Differentiation media hBMSCs differentiation into chondrogenic cells

Get tips on using Mesenchymal Stem Cell Chondrogenic Differentiation Medium to perform Stem cell Differentiation media hUMSCs differentiation into chondrogenic cells

Get tips on using MEGMTM Mammary Epithelial Cell Growth Medium BulletKitTM to perform 3D Cell Culture Media BT-549 cells-Mammospheres

Get tips on using Quant-iT™ RiboGreen™ RNA Assay Kit to perform RNA quantification Fuorimetric - human peripheral blood mononuclear cells (PBMCs)

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary porcine primary airway epithelial cell

Get tips on using ROS-ID® Total ROS detection kit to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

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.