... 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...

...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...

...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 (...

...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...

...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 ...

...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...

...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...

...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...

...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...