siRNA / RNAi /miRNA transfection Human Cells KG-1

Get tips on using PE Annexin V Apoptosis Detection Kit with 7-AAD to perform Apoptosis assay cell type - T-cells Mouse (OT-I)

Get tips on using HyClone Dulbecco's Modified Eagle Medium (DMEM) with high glucose: Liquid to perform 3D Cell Culture Media U87MG cells- glioblastoma spheres

Get tips on using HyClone Dulbecco's Modified Eagle Medium (DMEM) with low glucose: Liquid to perform Stem cell culture media hAdipose derived stem cells

Get tips on using Jump In™ T-REx™ HEK 293 Kit to perform Protein expression and purification Mammalian cells - HEK 293 HER2

Get tips on using pMT/BiP/V5-His A, B, & C Drosophila Expression Vectors to perform Protein expression and purification Insect cells - S2 HER2

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - BHK-21

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - mouse iPSC

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - mouse splenocytes

Get tips on using MEMα with L-Glutamine, Phenol Red, Sodium Pyruvate and Nucleosides to perform Stem cell Differentiation media mPericytes differentiation into Osteogenic 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.