dna-isolation-purification-yeast-pichia-pastoris

Get tips on using Chromous Genomic DNA isolation kit to perform DNA isolation / purification Bacteria - Gram positive Bacillus subtilis

Get tips on using MasterPure™ Complete DNA and RNA Purification Kit to perform DNA isolation / purification Cells - Immortalized cell lines C2C12

Get tips on using PolyATtract® mRNA Isolation Systems to perform RNA isolation / purification Yeast - Coprinus cinereus

Get tips on using PowerSoil® DNA isolation - DNeasy PowerSoil Pro Kit to perform DNA isolation / purification Bacteria - Gram negative Enterobacteriaceae

Get tips on using PowerSoil® DNA isolation - DNeasy PowerSoil Pro Kit to perform DNA isolation / purification Bacteria - Gram positive Mycobacterium tuberculosis

Get tips on using PowerSoil® DNA isolation - DNeasy PowerSoil Pro Kit to perform DNA isolation / purification Bacteria - Gram positive Lactobacillus amylovorus

Get tips on using Wizard® Plus Maxipreps DNA Purification System to perform Plasmid Isolation E. coli DH5α

Get tips on using DNA Isolation Kit for Cells and Tissues to perform DNA isolation / purification Cells - Primary cells Human primary keratinocytes

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 Wizard® Plus Midipreps DNA Purification System Technical Bulletin to perform Plasmid Isolation Proteus mirabilis