Protein expression and purification Mammalian cells

Get tips on using ApoAlert™ DNA Fragmentation Assay Kit to perform TUNEL assay cell type - Islets of langerhans (Beta cells)

Get tips on using ApoAlert™ DNA Fragmentation Assay Kit to perform TUNEL assay cell type - HeLa cells human cervical cancer

Get tips on using Gibco™ DMEM, high glucose, GlutaMAX™ Supplement to perform Stem cell culture media Mouse myoblasts cells

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb #12741 to perform Autophagy assay cell type - HK-2 cells

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb #12741 to perform Autophagy assay cell type - HK-2 cells

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb #12741 to perform Autophagy assay cell type - HK-2 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.

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 connexin 43 ShRNA to perform shRNA gene silencing Human - Neuroblastoma cells (SH-SY5Y) Connexin 43 lentiviral particles

Get tips on using HIF-1α siRNA (r) to perform siRNA / miRNA gene silencing Rat - Brain endothelial cells HIF-1α Lipid