Protein expression and purification Mammalian cells HeLa

Get tips on using ARCTURUS® PicoPure® DNA Extraction Kit to perform DNA isolation / purification Tissue - murine tail biopsies

Get tips on using Easy-Spin (DNA free) Total RNA Extraction Kit to perform RNA isolation / purification Tissue - Mouse Cornea

Get tips on using VWR Life Science RiboZol™ RNA Extraction Reagent to perform RNA isolation / purification Tissue - Human Gallbladder

Get tips on using PowerSoil® DNA isolation - DNeasy PowerSoil Pro Kit to perform DNA isolation / purification Yeast - Cryptococcus neoformans

Get tips on using miRNeasy Serum/Plasma Advanced Kit (50) to perform RNA isolation / purification Tissue - Livestock Blood / Serum / Plasma / Buffy coat

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 AllPrep DNA/RNA Micro Kit to perform RNA isolation / purification Tissue - Human Blood / Serum / Plasma / Buffy coat

Get tips on using PureZOL™ RNA Isolation Reagent to perform RNA isolation / purification Tissue - Rat Blood / Serum / Plasma / Buffy coat

Get tips on using QIAEX II Gel Extraction Kit (150) to perform DNA gel extraction / PCR product purification Product size > 15Kb

Get tips on using UltraClean 96 PCR Cleanup Kit (384) to perform DNA gel extraction / PCR product purification PCR clean-up