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If cloning methods had personalities, SLIC (sequence- and ligation-independent cloning) would be a true rebel. Not only does this system not use site-specific recombination, it also doesn’t require a ligation step. Based on the robust system of homologous recombination found in ...
If you’ve ever been looking for just the right CRISPR vectors on Addgene and found instead ones that were… pretty close, or at least close enough, you’ve found yourself with a common dilemma. Request the vectors you can find and use them as-is, saving time and effort but risking ...
All plasmids rely on their host cell's replication machinery in order to replicate—but not always to the same extent. As described in our previous Origin of Replication post, DNA replication is initiated at the ori and may or may not be synchronized with the replication of the ...
If you’re depositing a pooled library with Addgene, you may be surprised to learn that we ask for an amplification protocol with your deposit. This is because repeated amplifications of pooled libraries can lead to issues such as recombination and loss of plasmid diversity, so ...
Modular cloning is a popular DNA assembly tool used to build single- and multi-gene constructs for a variety of applications. MoClo tools can be used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even plants! In this blog we will review common ...
FITS In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However, in vitro screens cannot faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment that exists in vivo. Over the past five years, in vivo ...
Understanding the characteristics of natural plasmids and how they function in nature can inform us on how to create better recombinant vectors. In this blog post, we’ll define plasmid host range, identify a few of the features that confer broad host range in plasmids in nature, ...
