siRNA / miRNA gene silencing Rat Brain endothelial cells HIF-1α

Get tips on using pFastBac1-sccaIL12opt to perform Protein Expression Eukaryotic cells - Hi5 IL12 canine

Get tips on using GenJet™ In Vitro DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines MCF-7

Get tips on using Mesenchymal Stem Cell Osteogenic Differentiation Medium to perform Stem cell Differentiation media human umbilical mesenchymal stem cells (hUMSCs) differentiation into osteogenic cells

Get tips on using pcDNA3.1-MRP4-his6 to perform Protein Expression Eukaryotic cells - HEK293 human MRP4-his6

Get tips on using Gibco™DMEM, low glucose, pyruvate to perform Stem cell Differentiation media human umbilical mesenchymal stem cells (hUMSCs) differentiation into osteogenic cells

Get tips on using miRNeasy Serum/Plasma Kit to perform RNA isolation / purification Tissue - Human Blood / Serum / Plasma / Buffy coat

Get tips on using AmpFLSTR™ Identifiler™ Plus PCR Amplification Kit to perform Cell line authentication Human iPSC cells derived from peripheral blood mononuclear cells

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 Atg12 (D88H11) Rabbit mAb to perform Autophagy assay cell type - RPE-J cells

Get tips on using Beclin-1 (D40C5) Rabbit mAb to perform Autophagy assay cell type - K562 cells