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...you reach for that silica spin column, stop to consider some ways to purify DNA without a kit. DNA purification... kits have advantages: they are convenient and provide uniform, consistent results. But they are also ...method described in Kim and Morrison is nearly identical to that of commercially available kits but instead...concentration of a chaotropic salt (guanidine hydrochloride for the plasmid purification protocol and guanidine... Image: Kim and Morrison, 2009. This method provides DNA yield and quality similar to that obtained.... Homemade buffers performed the same as those provided with the kit. Although not tested by the authors... Molecular Biology Reference Visit our protocol video library References Kim, Y., & Morrison, S.L. (2009...

...biological disciplines and scientists employ GFP for a wide number of functions, including: tagging genes for... in the wildtype protein and facilitating its widespread use in research. Many other mutations have since... that many fluorescent proteins found on the red side of the spectrum are not GFP derivatives, but are...folding, maturation, and fluorescence. Biosensors: A wide array of GFP-based fluorescent biosensors has been...plasmids often include GFP as a marker to help identify which cells have successfully taken up the plasmid...applications for GFP. It has also been used to identify particular cell populations in drug screens, to...Plasmids 101 Articles Addgene's Fluorescent Protein Guide Note: A. Max Juchheim contributed to the writing...

...around $45,000, but as I’ve discussed elsewhere (slides here) there are many postdocs paid at much lower...must be met. In the Department of Labor’s general guide to the effect on higher education, you can see that...and Brandeis universities have stated that they consider postdocs paid on stipends to be FLSA exempt, and...institutions; and that institutions which can more easily provide bridge funding to absorb the cost will encounter...jobs that the majority of postdocs end up getting outside academia. There is also a valuable lesson for the...level they will be raised to on Dec 1st - across a wide consensus of academic stakeholders. There was ample...going away, and it is becoming ever clearer that outside scrutiny of science and the academic system is ...

...in an in-depth and easy-to-understand guided manner. These videos can be especially useful when designing...you begin, be sure to check out Addgene’s MoClo guide for all the basics of the process. Also refer to... The Sainsbury Laboratory Golden Gate Cloning videos made by the Nicola Patron group, one of the developers...and it will create the resulting plasmid map and identify any mismatches that are preventing your overhangs... 2 assemblies using the plasmid sequence files provided with every Addgene MoClo kit through their online...Additional resources on addgene.org Addgene's MoClo Guide and Kits MoClo Toolkit MoClo Plant Parts Kit ...

...hypoxanthine (shown) is the nucleobase component of the nucleoside inosine. Created with BioRender.com. Inosine...act on newly-deaminated inosines. Pro tip! The nucleoside inosine consists of the nucleobase hypoxanthine...transversion editor AYBEv3. This tool still retained considerable A → G editing activity at most sites, in addition...initial editor AXBE, for A → X base editor (X = any nucleotide). To create a more specific A → C base editor...be useful for saturation mutagenesis, and they provide a strong proof of concept for further development...efficiency, purity, and indel rate of each tool vary considerably. And there’s so much more exciting protein engineering...base editing Get a broad overview with our CRISPR Guide Check out our CRISPR 101 ebook ...

...site, and titer. Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult... bias: Addgene uses a prioritization process to decide which tools we make available. This prioritization...community. It does, however, necessitate us first identifying broadly useful vectors, a process that isn’t ...information from requestors and depositors, and identifying trends in our own repository data, giving us ...viral vector educational resources, from eBooks to guides to protocols, which was why I was so excited to...2023; Haery et al., 2019; McDade et al., 2016), video, educational resources, and collaborations, and ...Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using Pooled Lentiviral CRISPR Libraries...

...letters, while higher-level jobs are more likely to consider cover letters an important part of the application... content team, where writing is a core skill, considers cover letters to be very important. On the other...part of the job application process, it's a good idea to talk to people in the field to understand what...cover letter for? A cover letter is a place to provide helpful context about your candidacy, beyond what...make it easy to master ddPCR.  Finally, it’s a good idea to state why you’re applying to the specific company...unofficial poll, hiring managers also said that they considered cover letters to be more important for higher-level...Additional Resources Hamilton College Cover Letter Guide How to Write a Cover Letter Blog Post from Hubspot...

...invariably inconclusive and unreliable. Proper controls provide the constant variables that enable the correct ...functionality of your experimental system and help identify opportunities for troubleshooting or optimization...experiment, untreated cells should be included as these provide the baseline/standard against which other samples...correct interpretation of the results because it provides an important reference point when analyzing the... but the papers cited in the reference section provide more in depth information. Technical replicates... time consuming and ultimately more important. Ideally, each biological replicate should use fresh media...plasmids, be performed on different days, etc, but outside constraints may impose some limitations and you...

... of multiple animals on different occasions via video, generating huge amounts of data. When published...experiment would require access to each individual video. To PLOS’s credit, the organization subsequently...subsequently clarified what the policy intent was and provided an FAQ list to further explain the policy. The fact...variation is calculated and placed in the legend. Widely reported difficulties in reproducing experimental...the author or their host institution expected to provide data in formats exported to common file types? ...could be mitigated by on-line services such as SlideShare, Figshare and Dataverse. Just as Addgene helps...than just a single projection of it, could allow idea generation and extension, based on existing, published...

...with them. If you decided to make your own competent cells using the protocols provided above, then you ...cells are available online. A few examples are provided here for reference: Untergasser Lab Preparation...phase, then wash and resuspend the cells in the provided buffers. The competent cells are then aliquoted...Stbl3TM. How to transform your plasmid DNA If you decided to use commercial competent cells, then you're ...transforming your cells? Check out our transformation guide.   ...

...mutations found in leukemia, the Lowe lab was able to identify a set of mutations that made cancer cells more...mouse line. Over the years the fly has become an ideal model organism to study an array of topics including... to create transgenic animals. Zebrafish are an ideal model organisms for embryonic development studies... genes have an ortholog in C. elegans making it ideal to study basic molecular biology processes. C. elegans...quiescent and proliferative pancreatic β-cells identified by HOTcre mediated labeling." Proceedings of ...Menezes, Regina, et al. "From the baker to the bedside: yeast models of Parkinson's disease." Microbial...Additional resources on the Addgene blog Read our quick guide to Drosophila Download the fluorescent proteins ...

...Addgene’s catalog is a new class of sensors for neuropeptides and neurotransmitters. Motivated to develop ...GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost A to Z! - from ATP...vectors (here and here).  Figure 3: JEDI-2P was identified from a high-throughput screen, selected for increased...existing sensors. Using genome mining and structure-guided engineering of potassium binding proteins, they...point for further engineering. Figure 5: Structure-guided design and genome mining both led to new sensors...Fluorescent Potassium Indicators through Structure-Guided and Genome Mining Strategies. ACS Sens. 2022 May...protein plasmids and resources Fluorescent protein guide: Biosensors Collection ...

...loading control: the intensity of each POI band is divided by the intensity of the loading control in the ...problems for proteins of very different abundance, consider the following example gel. Figure 5: An...intensity axis. But the loading control points are outside of the linear detection range. As a result, they... problematic for computational quantification. Outside the linear detection range, band intensity measurements...experimental conditions. Otherwise, you might want to consider a different loading control. Assumption #2: The...final blot. Quantifying the entire lane of protein provides a loading control that can be used for normalization...The necessity of and strategies for improving confidence in the accuracy of western blots. Expert Review...

...pathway. “The idea is that if you introduce them into cells one at a time, you could identify those that ...apoptosis. For each of these 17 pathways, they identified a set of one to three mutant complementary DNAs... Wood says. “We want to make these available as widely as possible.” The researchers have plans to expand...References 1. Martz, Colin A., et al. "Systematic identification of signaling pathways with potential to confer...Lineage Tracing Read How CRISPR Can Be Used in Genome-Wide Screening Browse 28 Hot Plasmid Technologies from...

...New ideas and approaches are encouraged in science. It isn’t surprising that new ideas are also popping...before you even receive the product? How can we consider the environmental impact of those processes? Who...might sound risky in terms of contamination, but consider flow cytometers. These fluidic machines handle...through an EPA-approved process and no longer considered hazardous. Then, they use this material to produce...plastic lumber and distribute it to industrial and residential markets. Rethinking our approach to lab waste...

...Accessibility Guidelines (WCAG). These changes mean our content is both available and accessible to a wider community... Over 150,000 plasmids empowering researchers worldwide. Countless breakthroughs enabled by shared resources...part of the story. This year, we made significant strides in making science more accessible and reliable....community. As we enter 2025, your feedback continues to guide our innovation.  In response to a demand for more...rapidly, and with it, the needs of researchers worldwide. Through our growing community on Bluesky, our...

...How to put the wider scientific community at the service of COVID-19 research? We decided to give it a ...internationally COVID-19 Resources Canada is a database that provides information about reagents, funding and volunteer... the information is applicable to researchers worldwide. “COVID-19 has spread incredibly fast and without...can advance COVID-19 research to hopefully help guide policies and ultimately treatments in a timely fashion...Read about COVID-19 research on the blog Visit our guide for external collaborations Resources on Addgene.org...

...proteoglycan (HSPG)3 AAV receptor (AAVR)2 Epidermal growth factor receptor (EGFR)3 AAV8 Laminin...there are up to thirteen natural serotypes of AAV identified so far, numbered AAV1, AAV2, and so on. In addition... the second number to the capsid. Most vectors provided by Addgene are pseudotypes using the genomic structure...AAVrg has not been thoroughly studied in contexts outside the nervous system to confirm its cellular receptors...addition to the toolbox, with the first paper identifying a binding receptor published this year (Dhungel...2008). Adeno-Associated Virus (AAV) Serotype 9 Provides Global Cardiac Gene Transfer Superior to AAV1,...Systemic Capsids Resources on Addgene.org AAV Guide AAV Data Hub AAV Packaged On Request Browse all ...

...every project we undertake and every service we provide is tested against fulfilling our mission. “Addgene...continuously improve all the extra services we provide like quality control, curation, technical service...starting new initiatives as opposed to fundraising.   Aside from enabling us to make all decisions with the ... keeping excellent records and maintaining confidentiality where it is appropriate. This grows our relationship...Addgene increases productivity by providing world-wide access to curated, high-quality data and reagents...processes, distribution, and customer service) Addgene provides unmatched customer service that is consistently...

...middle of the targeted region, as shown below.  To identify optimal nickase designs, IDT scientists Mollie...insert an EcoRI site using a single-stranded oligonucleotide (ssODN) donor. Using a D10A nickase to insert...nickase outperformed both WT Cas9 sites tested by a wide margin, exhibiting >20% repair efficiency. Clearly... is especially useful if there are no available guides close to the desired mutation site. Quick tips...small insertions or tags (ssODN; IDT Ultramer® Oligonucleotides) Use 100 nt homology arms for large insertions... Find CRISPR Nickase Plasmids Browse the CRISPR Guide ...