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This post was contributed by guest blogger, Krissy Lyon, a PhD candidate in Neuroscience at Harvard University. Just as computers, cell phones, and cars become more technologically advanced leaving earlier versions obsolete, the techniques we use in lab are replaced by improved ...
This post was updated on March 21, 2018. Most of the time, plasmid prepping is a breeze. You get your stab from Addgene, streak for single colonies, sub-culture, and prep with a DNA prep kit or your lab's favorite in-house protocol. DNA yields for this procedure are typically in ...
Molecular cloning requires some method of screening colonies for the presence of an insert. Traditionally this has been done with restriction enzyme digest; however colony PCR can accomplish the same thing in less time and for less money. The key steps to colony PCR are: 1) ...
Two hybrid systems were developed in Saccharomyces cerevisiae in 1989 and are still used extensively to screen for molecular interactions in the cell, including protein-protein, protein-DNA and protein-RNA interactions. The 1980s saw a flurry of discovery in the field of ...
Congratulations, you have a plasmid expressing your gene of interest (YGOI) and are ready to dive into your functional experiments! Whether you’ve cloned the plasmid yourself or obtained it from a colleague down the hall, it is always a good idea to take some time to confirm ...
This blog was originally published on BitesizeBio here. As part of my job ensuring plasmid quality at Addgene, I analyze 50-100 sequencing reactions a week. So I have developed some good habits that I wanted to pass on to you to make sure you are getting the most out of the data ...
This post was updated on Dec 4, 2017. At Addgene, we continually use the Basic Local Alignment Search Tool (BLAST) provided by NCBI. BLAST helps us compare the sequencing results of the plasmids in our repository with known reference sequences, such as full plasmid sequences ...
