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last updated: July 28, 2023
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Struggling with your cloning? It could be time to try blunt-end cloning. This easy introduction explains what it is, when to use it, and how.
Isolating pure DNA is key to many downstream applications for molecular biologists. Isolating large quantities of pure DNA used to be a laborious task. But thanks to commercially available kits, older methods have been streamlined to allow efficient recovery of pure DNA. In this article, I will talk about a method called DNA gel extraction,…
What Is Alternative Polyadenylation? Processing of mRNA and its regulation plays a fundamental role in gene expression. As science progresses, alternative polyadenylation takes center stage in the undercurrents of gene expression. 1,2 Polyadenylation is part of the pre-mRNA maturation process and involves polyadenylation of the 3’ end of the emerging RNA. This process happens to…
Most of us are aware of genetic engineering systems like Cre-Lox, TALENs, Zinc finger systems, and of course, CRISPR-Cas9. These are all examples of CSSR- Conservative Site-Specific Recombination. We use these site specific recombinases routinely, but do we really know about them or what the future hold for these tools? It turns out that CSSR…
In my last article, I explained that plasmid DNA recovered from a plasmid prep consists of few different species; supercoiled, nicked, linear and single stranded circular, and how you can distinguish them on a gel. Supercoiled DNA is the desired form of plasmid DNA; it performs better in downstream applications such as automated sequencing and…
In Part I of AAV Production, I described how to produce crude (non-purified) AAV. In this article, I am going to tell you how to purify that crude prep. Virus purification is usually done by gradient ultracentrifugation. Two common methods involve gradients made from increasing concentrations of cesium chloride or iodixanol. A cesium chloride prep…
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