...) and are ready to dive into your functional experiments! Whether you’ve cloned the plasmid yourself or...determine the relative sizes of the resulting DNA fragments. Restriction digests are commonly used to confirm...longer to achieve good separation of larger DNA fragments. I hope these tips demonstrate that plasmid ...

...the N and C terminal segments respectively of a GFP dependent Cre. The two segments come together in the...transduction patterns empirically for their own experiments. To this end, we offer control AAV in smaller...

...skills, while also learning about new scientific developments and helping scientists is a win-win for me. ...shared in the post. And the blog writing tips are elements I try to include in each post I write for the ...your research or career? Share your story in the comments below!   Additional resources on the Addgene...

... and you’ll need to track it throughout your experiments.  Now that that’s out of the way, you can put... reaction to understand if any of your buffer components could affect the reaction. For instance, sodium... - antibodies that are a good match for your experiments, and avoid the awkwardness of getting the right...

...they successfully imaged multiple subcellular compartments over several minutes using Nano-lantern fusion...single molecule imaging. Nano-lanterns and your experiments The Nagai laboratory has a done a wonderful job...Addgene. Suzuki, Kazushi, et al. “Five colour variants of bright luminescent protein for real-time multicolour...

...being depicted, we can’t forget that those other components are still there in real life.  Certainly referring...specifically study it. They might be researching enhancer elements that control the transcription of a gene a megabase...versatile, small, sensitive immunohistochemical reagents. Yamagata M, Sanes JR Proc Natl Acad Sci U S A...

...idea that would evolve into Sample6 as graduate students in Dr. Jim Collin’s Lab in 2009. From 2009 to ... an idea or product? Please let us know in the comments section! And read on for more great information...contributed by Julia Phillips and Fiona Beckman Acknowledgments Thank you to Dr. Mike Koeris (Co-founder and...

...such as RNaseZAP. When harvesting tissues, cells, plants, fungi, or bacteria, keep samples cold and work...the homogenate was loaded onto cesium-chloride gradients and ultracentrifuged for up to 20 hours to separate...instead of RNA separation using cesium-chloride gradients, water-saturated phenol, sodium acetate, and chloroform...

...restriction sites to easily assemble multiple DNA fragments. This system has been used to construct plasmids...McCormick lab generated knock-out and knock-in mutants, determined protein expression levels from different...critical for reducing Cre toxicity in long-term experiments.  Figure 4: The C-terminus of Cre (CreC...

...about it. Building a diverse team with different talents and expertise Fortunately, the eLife ambassadors... had previous experience organizing open access events and grassroots efforts. After thoughtful conversations...Twitter (@repro4everyone), or email the team. Acknowledgements One of the highlights of 2018 for me was to...

...Addgene, so troubleshooting molecular biology experiments was in the job description, but this skill is...broader research questions and designing future experiments. Being able to work through these types of bigger....     Know your strengths! Research experience presents ample opportunities to practice problem solving...

...CRISPR-Cas systems SEND plasmids Prime editing improvements Miniature CRISPR-Cas systems By Brook Pyhtila... from Segel et al. 2021.   Prime editing improvements By Angela Holmes Several labs have recently ...PEDAR is capable of correcting larger genomic rearrangements as demonstrated in a mouse model of tyrosinemia...

...illumination in in vivo, ex vivo, or post-mortem experiments. Fluorescent proteins can also be identified ...transgenes of interest. Furthermore, fluorophore variants (for example green and red fluorescent proteins...and could even be beneficial in some types of experiments (for instance, when the fluorescence expressed...

... 4B and C in Zhou et al for details on these experiments. Photoswitchable kinases could possibly be used...can I try using photoswitchable kinases in my experiments? You can find the plasmids from Zhou et al here...Carrasco, and Markus W. Covert. "High-Sensitivity Measurements of Multiple Kinase Activities in Live Single...

... aggregate. The basics and pitfalls of these experiments are discussed here. In vitro tests for aggregation...your cell type of interest before starting your experiments.  Selecting a non-oligomerizing protein A fluorescent... Of course, you want the ‘latest and greatest’ variants with the best properties for your research. But...

...reactions on tissues can be cumbersome Tyramide reagents can be expensive Can only stain with one HRP-RANbody...consider leaving a link to its sequence in the comments below. References Sanes, J.R., & Yamagata, M....versatile, small, sensitive immunohistochemical reagents. PNAS. PubMed PMID: 29440485. Additional Resources...

...wash steps removes excess proteins and other contaminants. The protein of interest is then eluted using...polyhistidine-metal ion interaction is not disrupted by denaturants (Hochuli, 1988). In some cases, proteins can ...more stringent wash conditions and remove more contaminants without losing your protein of interest. But...

...It’s also a great way to keep track of past accomplishments. Follow all the rules of good resume writing...writing. Include examples of concrete accomplishments in both the technical and “plus skills” areas. Include...’s news and updates. Share your own good ideas, events and favorite resources. Respect the network and...

...introduction, methods, results, conclusions, and acknowledgements sections. Some other additions you might wish...present at your poster at all times, so other participants can identify you. You may also wish to make ...the introduction, methods, conclusions, and acknowledgements sections are inserted. The advantage of this...

...of the mutations engineered into each of their parents. Here’s how to cross mice, using a simple example... all of your desired alleles to conduct your experiments. You should also maintain your parental strains...contain a mix of genetic material from each of the parents. If both of the mutations that you are trying to...