To avoid cross-contamination in qPCR, key practices include: separating pre- and post-amplification areas with dedicated equipment, using a unidirectional workflow, including a “no template control” in each run, properly cleaning work […]
Miscellaneous
Reduce the risk of cross-contamination from the amplicon/template and previous reactions for a qPCR
Use a unidirectional workflow during experiment setup. Separating pre- and post-amplification areas is key to preventing contamination. Prepare your PCR master mix in a template-free room (see next bullet) using reagents that never […]
Difference between dye-based vs probe-based qPCR
Dye-based qPCR is less expensive than probe-based qPCR, but probe-based qPCR is more specific and accurate. The choice between the two depends on the research goals and the type of […]
OmpA Signal for Protein Expression
The OmpA signal peptide is a 21-amino acid sequence that targets fused proteins to the Sec secretion pathway in E. coli. It can be used to increase protein yield and simplify purification […]
High-cell-density cultivation (HCDC)
High-cell-density cultivation (HCDC) is a process that improves the formation of microbial biomass and products. It can be used for microorganisms such as bacteria, archaea, and yeasts. HCDC can make the fermentation process […]
How to protect RNA from degradation?
To protect RNA from degradation, you can: RNA is prone to enzymatic degradation and hydrolysis. Even at low temperatures, RNA can still be reactive, and some ribozymes can be enhanced by […]
What is the main reason for RNA being unstable?
Unlike DNA, RNA is usually single-stranded. Additionally, RNA contains ribose sugars rather than deoxyribose sugars, which makes RNA more unstable and more prone to degradation. RNA is generally stable at –80° […]
Potential Reasons for Lack of Supercoiled DNA
Conclusion In conclusion, the absence of apparent supercoiling in plasmid DNA preparations can result from a variety of factors related to DNA extraction, purification, handling, and analysis. By systematically addressing […]
Transfection Efficiency: What Makes Plasmid DNA Transfection Grade
Successful transient transfection relies on many factors, starting with a successful plasmid prep. To ensure high transfection efficiency, resulting DNA from your plasmid preps must be transfection grade. But what […]
Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands
When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA […]