DNA isolation / purification Plants

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.

DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

Get tips on using miRCURY Exosome Cell/Urine/CSF Kit to perform Purification of extracellular vesicles Exosomes - Seminal plasma

Get tips on using pOPINE to perform Protein expression and purification Bacteria - Escherichia coli medin

Get tips on using QuickLyse Miniprep Kit (250) to perform Plasmid Isolation Enterobacteriaceae

Get tips on using QIAGEN Plasmid Kits to perform Plasmid Isolation E. coli DH5α

Get tips on using GeneJET Plasmid Miniprep Kit to perform Plasmid Isolation Piscirickettsia salmonis

Get tips on using GeneJET Plasmid Miniprep Kit to perform Plasmid Isolation Proteus mirabilis

Get tips on using CompactPrep Plasmid Midi Kit to perform Plasmid Isolation Yersinia pestis