...bloggers Becky Kucera, M.Sc. and Eric Cantor, Ph.D. from New England Biolabs. Becky Kucera is a Principal...This post was contributed by guest bloggers Becky Kucera, M.Sc. and Eric Cantor, Ph.D. from New England ... more than 10 modules often rely on two-step hierarchical approaches using different Type IIS restriction...

...that allowed researchers to create stunning, interactive images of their proteins. Since the 1980’s, journal...obscures the flexible nature of all proteins in general, and the amazing conformational flexibility that...strand. In addition, this model also displays the several Lys/Ala mutations reported by Ian Slaymaker and...classroom or trying to explain your science to the general public, visit the Center for BioMolecular Modeling...

...trail at the top of any plasmid page gives the hierarchy of web pages the plasmid page is nested in. You...it? Your best bet is the Plasmid Map: We auto-generate a plasmid map based off of the plasmid sequence...plasmids are sent as bacterial stabs at room temperature. Core of the plasmid page - Core of the plasmid...), antibiotic resistance, and optimal growth temperature are listed, as well as the name of the strain...

...polyclonal antibodies are readily available from several sources, fewer sources of recombinant antibodies...appropriate buffer, it’s time to quantify it. There are several options to choose from depending on the time, reagents...2018). Aggregation can arise from a shift in temperature, pH, or salt concentration (Amin et. al., 2014... shipped to customers on ice, preventing any temperature shifts. Of course, we are also cautious during...

...set up a standard PCR reaction (primers, dNTPs, polymerase) using the supernatant of lysed bacteria as template...standard PCR reaction: combine template, primers, polymerase, and dNTPs and then incubate with a standard ...heating step of the PCR reaction. A standard Taq polymerase is sufficient. Controls: Controls can make ...them onto the gel. The figure above summarizes generalized expected results for the three primers previously...

...multi-drug resistance: definitions vary in the literature, but generally organisms with multi-drug resistance are...abstract, convoluted scientific concept. It is imperative that we all understand what antibiotic resistance...meats. We can focus on creating new antibiotic therapies, and investigate more effective drug targets (...

...multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating purification...labeling, enrichment on streptavidin and next-generation sequencing). Using this sensitive method, they...that the field has overcome with split AAVs. In general split AAVs break a large transgene in two pieces...  Wang D, Zhang F, Gao G (2020) CRISPR-Based Therapeutic Genome Editing: Strategies and In Vivo Delivery...

... are several different versions of the plasmid partitioning system in the bacteria, but generally it consists...coresident plasmid. Asymmetric plasmid loss occurs for several reasons, including one plasmid blocking replication...compatible or not.  Using plasmid incompatibility for therapeutic advantage Although it may seem like plasmid incompatibility...

...quantitative imaging experiments and cell physiology in general. Pitfalls of Fluorescent Protein Fusions Observing...Previously, we have described a number of practical considerations that should be taken into account when deciding...cellular compartments other than the cytoplasm. Generally, FPs evolved in or were engineered for use in ...N-glycosylation consensus sequences in popular FPs. This generated a palette of inert green, cyan, yellow, and blue...

...efficient method to generate precise, unmarked mutants, but it frequently requires considerable optimization ...transformation and plasmid curing steps, and can generate off-target mutations. Modifying bacterial genomes...clinical infectious diseases at the Massachusetts General Hospital where he is now an attending physician...

...different size and structural complexity. After several steps of tissue processing prior to running my ...voltage in the cytometer’s software in order to generate signals in a reasonable range. If colleagues in... being considered for the analysis. (C) A more liberal gate allows more cells to be considered for the...some like to make looser ones (Figure 3). A more “liberal” gate in the beginning should not have a great ...

...of understanding how the body works, generating new therapeutics, and sometimes formulating new biological...Bio-protocol all work to provide more descriptive and interactive protocols that make it much easier to learn new...Initial positive results and the excitement they generate can, however, bias us. We’ve all heard stories...

...binds the plasmid to the new fragment thereby generating a complete, circular plasmid that can be easily...green, insert) are cut with restriction enzymes to generate compatible overhangs that allow them to bind. ...single tube allowing the compatible overhangs generated by restriction digestion to anneal to one another...midiprep or a maxiprep. In these purifications, you generally lyse the bacteria; add chemicals to precipitate...

...base editing from the first generation of base editors to the eighth generation (and dual base editors, too...been updating many of these posts over the last several months. Now the #12DaysOfCRISPR is back on Twitter...

...process, simply enter your preprint title or a general description of the deposit. Select your desired...millions. Pooled libraries must be verified by next-generation sequencing. Please see the help center article...that can be used under BSL1 conditions were generated in your lab are necessary for use of a plasmid...need information on how to verify any genomic alterations unique to your strain. Please contact us at deposit...

...Kit from John Chodera's Lab - Get high expression of His-tagged human kinases in bactera. Listen to Our...Listen to the entire Hot Plasmids episode! We generally publish 5-10 Hot Plasmid articles with each of...

...a Clinical Fellow in Pathology at Masachusetts General Hospital and Postdoctoral Fellow in the Church ...gene drives focusing on safety and the ability to generate mutant microbial strains faster than ever before... Advisory Board with a particular interest in generating Cas9 based tools to enable facile control of ...

...service to further contribute to our mission to “accelerate research and discovery.” Here are the details...in the research on the SARS-CoV-2. Nine of the overall plasmid selection will be open to industry scientists...preparation is quality controlled through next generation sequencing and spectrophotometry to test purity...

...researchers facilitate and accelerate scientific discovery. How can Addgene help accelerate your research? Let...Addgene headquarters every day. That would be an average of 673 plasmids each day. This requires tremendous...

...dual-nuclease Cas9-Cas9 chimeras Scot Wolfe’s lab has developed dual-nuclease Cas9-Cas9 chimeras with higher target... in Nature Communications Find the Cas9-Cas9 chimera plasmids A modular toolkit for genome editing...