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...researchers to identify and isolate very specific biological targets. However, as popular and useful antibodies...antibodies, creating a potentially unlimited supply of identical antibodies (See Fig 1.). rAbs are a relatively...has specific binding and has a strong signal. We call the unique pattern of the antibody binding “distinct...TS (2015) The Economics of Reproducibility in Preclinical Research. PLoS Biol 13(6): e1002165. https://...

...choosing antibodies   Complexity is a key factor Biological systems tend to be complex with a variety of ...multiple targets in parallel. In these cases, it is critical that users choose antibodies that are easily distinguishable...staining multiple targets indirectly, you must strategically plan primary/secondary antibody pairing such...Fisher, S. K. (1993). Chapter 15 Postembedding Immunocytochemical Techniques for Light and Electron Microscopy...

...plasmids. These are prone to error — both human and biological. Even the most high-fidelity polymerases can ...the correct inserts, tags, promoters, and other critical functional components. Fun fact! Some older plasmids...sequences. These sequences can also be confirmed or theoretical. No matter the level of information that is provided...the raw NGS reads and reassemble using a program called Geneious. Here, the raw reads are mapped against...

...things more interesting, we are working in a new technical discipline that nobody understands (synthetic ...thing is opening a pizza joint. Silly ideas (mining local salt deposits to sell at farmers markets), and serious...that survived the literature review, survived technical feasibility analysis, and was a marketable product...and rapidly indicate their presence. They are genetically modified organisms, and GMOs have a bad reputation...

...can be long—5 years or more, especially in the biological sciences—whereas industry postdocs have a set...course, you might have less time to explore scientifically since you can’t afford to spend too much time...Unless your academic lab is closely involved with clinical trials or has ties to industry, the drug discovery...industry postdoc, you might find yourself feeling scientifically isolated if no other postdocs are in your department...

...margins of my protocol. While these tips weren’t critical to the experiment, they definitely made my life...qPCR Protocol for more details. Pro Tips When calculating the titer for your AAV, remember to account for...control (NTC) should always be included. While the physical titer generated by qPCR is useful, I test a few...tool for gene expression in mammals Use the PEI calculator for planning AAV packaging transfections Find...

... Liu, 2023). This method uses a prime editor (typically a Cas9 H840A nickase fused to a reverse transcriptase... design of the pegRNA has been shown to play a critical role in the editing efficiency of prime editing...experimentalist, and especially shines once large-scale prime editing experiments come into play. Nicolas...s41587-023-01678-y Li, X., Chen, W., Martin, B. K., Calderon, D., Lee, C., Choi, J., Chardon, F. M., McDiarmid...

...therapeutic strategies to target it, than you typically would at a microbiology meeting. Create an outline...lab mates, and PI can provide feedback on your technical presentation, but those outside of your field ... through the list at the session so you can strategically use your time. Don’t have access to a list of...titles to find the ones you’d like to revisit. Strategically traverse the sea of posters If you come prepared...

...the ωRNA (TnpB) or a gRNA (Cas9). Drawings not to scale. Created with BioRender.com. Steven Jacobsen...Rodriguez-Fraticelli recently developed an approach called Simultaneous Tracking of Recombinase Activation...recombinase payload activates a floxed reporter gene (genetically encoded or delivered separately). Figure...Cre-dependently to allow tracing exclusively from genetically determined cells. This version of ATLAS works...

...your gene of interest is difficult to target specifically with a gRNA). Be sure to keep these techniques...sequences are exchanged between two similar or identical molecules of DNA. Gene targeting takes advantage...homology is required, but 6 to 14 kb of homology is typical for targeting constructs. In the example shown ...cells participating in development or other physiopathological events to which the chosen promoter responds...

... 2017. At Addgene, we continually use the Basic Local Alignment Search Tool (BLAST) provided by NCBI. ...BLAST sequence databases from the dropdown menu. Typically, we use the default nucleotide database “Nucleotide...” program for blastn so that you can extract practically every single reliable base pair from your results... the “Align two or more sequences” option to drastically reduce waiting time for search results. Optimizing...

...-length viral genome (Figure 2). While it is technically possible to encode all four viral genes in the... (see Uses of Minigenome System). From a more technical standpoint, a plasmid with all four genes and ... gene by altering the amount transfected (see Technical Tips). Minigenome replication and transcription...compounds which often target the viral polymerase. Technical tips It is important that all of these plasmids...

...lab tool. In the classic two hybrid system (also called the interaction trap), genes encoding proteins ...experiments. False positives can occur even in small scale experiments, although these can often be accounted...that the configuration of your fusion proteins physically blocks the binding sites for the protein partner...Tachihara, K., Umezawa, Y.l. Split luciferase as an optical probe for detecting protein-protein interactions...

...that, they also help us keep track of the tubes’ physical location in the lab. These barcodes additionally...  freezers depending on the sample type. It is critical for us to know which plate, rack, and freezer ...performs an average of 140 transformations. The lab typically transforms newly deposited samples from Monday...Excel sheet.3. Be specific  - When assigning a physical location to your samples, include as much detail...

...been working on development of technologies for biochemical analysis of genome functions such as transcription...researchers to use enChIP in a wide variety of biological systems. We are currently developing several ...Therefore, my lab is dedicated to development of new biological technologies. The locus-specific ChIP technologies... effort to interact with each other, which is critically important, especially for young researchers. ...

... defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a single base...precisely changes a single base with an efficiency typically ranging from 25-75%, while the success of precise... in the DNA by cutting only a single strand, dramatically reducing the likelihood of detrimental indel...known 'DNA adenosine deaminases' which would theoretically give rise to an adenine base editor. A massive...

...passed down through generations (Figure 1). These so called “plasmid addiction systems” require dividing cells..., plasmid addiction systems have been used as biological tools for a wide range of applications in molecular...become a powerful tool to address a diverse set of biological inquiries. References Ogura, Teru, and Sota...vaccine strain CVD 103-HgR." FEMS Immunology & Medical Microbiology 57.1 (2009): 69-79. PubMed PMID: 19678844...

... ligands (Figure 2). Biosensors, which include calcium and voltage sensors, are the broadest category ...catalog If you’ve been following our blog, you may recall our first 20th anniversary story on CRISPR, where... vectors available in the repository. GCaMP, a calcium indicator (biosensor, by Addgene’s categorization...Waxmonsky, N. C., Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using Pooled Lentiviral CRISPR...

...studying parenting strategies and make important ecological indicators since species live in water and on... as the only Monarch butterfly food. They have chemical pathways generating interesting drug-like compounds...Lab (@EvoEmbryo) September 24, 2018   Xenopus tropicalis (Gary Gorbsky) This is not a new experimental...plants and are unusual enough to be interesting biologically. They have economic importance as harmful weeds...

...gives Snapgene great exposure to the community of biological scientists who use Addgene everyday. To get this...from other institutions personally on conference calls and make sure to maintain these relationships by...speakers for a meetup or recruit volunteers for your local animal shelter. There are many opportunities so ...Universities themselves can help you do this through practical tools like alumni networks. I find it useful to...