shRNA gene silencing Mouse Prostate cancer cell lines (DU145 and PC3)

Get tips on using Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham to perform Mammalian cell culture media T47D

Get tips on using The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit to perform Autophagy assay cell type - T98

Get tips on using The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit to perform Autophagy assay cell type - A375

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 HyClone Dulbecco's Modified Eagle Medium (DMEM)/ F12 1:1: Liquid to perform Stem cell Differentiation media hDPSCs differentiation into adipogenic cells

Get tips on using STEMdiff™ Hematopoietic Kit to perform Stem cell Differentiation media hiPSCs differentiation into CD43+ primitive hematopoietic progenitor cells (HPCs)

Get tips on using Gibco™ StemPro™-34 SFM to perform Stem cell culture media Mice spermatogonial stem cells (mSSCs)(C18-4)

Get tips on using Gibco™Essential 8™ Medium to perform Stem cell Differentiation media hPSC or hiPSCS differentiation into myogenic cells

Get tips on using TACS® 2 TdT Fluorescein Kit to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells