siRNA / RNAi /miRNA transfection Human Cells THP-1

Get tips on using DC™ Protein Assay Kit I to perform Protein quantification Mammalian cells - HeLa

Get tips on using Pierce™ BCA Protein Assay Kit to perform Protein quantification Mammalian cells - SiHa

Get tips on using Gibco™DMEM/F-12, no glutamine to perform Stem cell Differentiation media hPSCs or iPSCs differentiation into Lung progenitor cells

Get tips on using Gibco™ DMEM/F-12, GlutaMAX™ supplement to perform Stem cell Differentiation media iPSCs or hESCs differentiation into Neuronal cells

Get tips on using CellTiter-Glo® Luminescent Cell Viability Assay to perform Live / Dead assay mammalian cells - BHK-21

Get tips on using SV 96 Total RNA Isolation System to perform RNA isolation / purification Cells - primary porcine primary airway epithelial cell

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.

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 Cell DNA Isolation Kit to perform DNA isolation / purification Cells - Immortalized cell lines C2C12

Get tips on using Cell Counting Kit-8 to perform Live / Dead assay mammalian cells - rat nucleus pulposus