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...spent this remaining time by elaborately pranking each other, dressing in costume, and holding competitions... we are sending hundreds of plasmid requests out each day. As you may imagine, it keeps all of us quite...furthering our mission by hard work, we genuinely enjoy each other’s ideas and company. The Addgene summer picnic...sometimes work is a bummer when you’d rather be at the beach or binging a new Netflix series. The thing that ...you can enjoy the people you spend 40+ hours with each week. In conclusion - get out there any have some...

...number of days and tracking average transit time for each courier we contracted with so we can always offer...can see our most popular educational resources for each type of resource we offer.  In 2013, we launched...experimental help. We love it when a researcher reaches out to ask if they can help us create or update... We’ve learned that increased accessibility is achieved through interconnected incremental changes. It... Rachel Leeson...

...Tracking cellular history with protein ticker tape Teaching an old antibody new tricks   New compact, efficient...REcipient Cells Through Envelope Design). This modular approach combines an interchangeable set of fusogenic and...side effects. This platform offers a promising approach for experimental therapies and medical applications... into the TurboID enzyme, Lee and Cheah et al. achieved precise control over when and where the enzyme... bioRxiv — doi: 10.1101/2021.10.13.463862.    Teaching an old antibody new tricks By Ashley Waldron While...

...by Kusumika (Kushi) Mukherjee, a Postdoc at Massachussetts General Hospital. A little over a decade back...non-integrating RNA virus Episomal vectors- extrachromosomally maintained DNA plasmids that express reprogramming...encoding reprogramming factors behave as separate extrachromosomal elements in the nucleus of the target cell...Epstein-Barr Nuclear Antigen-1 (oriP-EBNA-1) system where each vector contains a viral origin of DNA replication...Mukherjee is currently a postdoctoral fellow at Massachusetts General Hospital with research interests in ...

...This post was contributed by ATCC Outreach Scientist Nick Amiss. Cell culture in the present day Cell... number, cell line, and date, which you complete each time you passage the cells (as well as recording...liquid nitrogen for your Working and Master stocks of each cell line. Once all Master vials have been thawed...the results are reliable. With this two-pronged approach, the many mistakes of the past can be prevented...desired clinical endpoints.   Nick Amiss is an ATCC Outreach Scientist at LGC Group, London, which is the European...

...anneal with each other, while the 5’ flaps containing the original DNA sequence anneal with each other. Excision...identify dozens or hundreds of particles surrounding each of the highly-visible beads on the cryo-EM grid.... samples during cryo-EM grid plunge freezing By Rachel Leeson Cryo-EM samples can be damaged at the air-water...plunge freezing represents a convenient, effective approach to protecting your cryo-EM samples from damage...

...expression, and viability. Gates are the parameters the machine uses to differentiate between variations in those...tumor cells, immune cells, blood cells and more), each coming with a different size and structural complexity...A) A flow plot showing two stains in relation to each other. Using a quadrant gate, I can divide my cell...cell population into DN, DP and SP (for each stain respectively). (B) Instead of a quadrant gate, I am...occur when a cell is pulsed by the laser. Within each type of scattering, you also discovered that there...

... will overcome certain obstacles, alternative approaches, potential collaborations, critiques of your ...and all of the caveats and alternatives for an approach, if asked. The key to a chalk talk is to effectively... the knowledge gap in the field? How will your approach resolve this problem? (2-3 sentences) What.... Identify the key points you want to convey for each section or aim. Make sure those get priority. The... fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance runner, gardener...

...obvious that a desired career outcome is not easily achieved. Of note, it was highlighted during the conference...Positive Points for Trainees - Taking Steps to Achieve a Successful Career Career Advice from the Career...showing researchers that they can take steps to achieve success in their careers. The take-away message...can take the time to do this, you should always reach out to your peers and other mentors who can help... postdocs, or professors isn’t nearly enough to reach diversity goals. Universities need to show that ...

...I have a list of what I really need in order to reach my research goals. Would I be able to get access...lack of resources, then ask for suggestions to meet each others’ needs and limitations. Everything is negotiable...research training to help you find other solutions to reach your goals. For example, a junior investigator turned...seem, you should consider it if it will help you achieve your research goals, even if that means saying ...a laboratory research management consultant and coach in Boston, MA. He specializes in assisting research...

...), but any HEK293T cell line should work. This approach can be adapted for different cell lines and different...ratio will need to be empirically determined for each new batch of 1 mg/mL PEI prepared. There may be ... used, volumes, pH adjustment etc. Consequently, each batch needs to be validated and the best ratio of...

...cell/DNA mixture on ice for 20-30 mins. Heat shock each transformation tube by placing the bottom 1/2 to...enough colonies for your next step. Remember that each of these shortcuts will reduce the efficiency of...cuvettes. Follow the manufacturer's instructions for each. I got no transformants. What went wrong? Check ...

...Place a sterile poly-D-lysine coated coverslip in each well of a 24-well cell culture treated plate. Seed... aspirate the media from the 24-well plate. Wash each well with 500 µL of PBS, remove the wash, and dispose...dispose of it in an appropriate waste container. Fix each well with 500 µL of cold 4% paraformaldehyde in ...

...happening everyday, it’s not possible for us to reach everyone. Therefore, to make our library more useful...the repository? Let us know! You can notify us to reach out to potential depositors in two ways: 1. Fill...include any additional details that will help us reach out to the corresponding potential depositors. ...plasmid. While we cannot guarantee that everyone we reach out to will end up depositing, we hope this campaign...

...histone modification at specified loci. Charles Gersbach’s lab has deposited mammalian expression constructs...Both of these methods also recruit epigenetic machinery - but is there an advantage to using direct epigenetic...Vockley CM, Thakore PI, Crawford GE, Reddy TE, Gersbach CA (2015) Epigenome editing by a CRISPR-Cas9-based...Safi A, Song L, Hilton IB, Crawford GE, Reddy TE, Gersbach CA (2017) CRISPR–Cas9 epigenome editing enables...Ruan K, Wang F, Xu G, Hu R (2016) A CRISPR-based approach for targeted DNA demethylation. Cell Discovery...

...our bloggers each and every month, we've decided to start an "Editor's Choice" series. Each month, I'll...was a fantastic month for the Addgene blog. We achieved roughly 50,000 views over the entire blog and ... Expression in Mammals Recombinase-based State Machines Enable Order-dependent Logic in vivo As always...

...related LCR technique uses two sets of probes to achieve exponential amplification. What does fidelity mean...substrates. Thus, high fidelity ligation can be achieved through a combination of the intrinsic selectivity...four upstream probes and four down-stream probes, each differing only in the base at the ligation junction...represented. Sixteen separate pools were prepared, each with a different complement strand covering all ...the template. The probes were designed such that each possible pairing resulted in a product of a different...

... used in limited circumstances. An alternative approach is to flank the desired change with 4-5 silent...from mini-λ or 4) from the lambda red phage itself (each of these will be discussed in more detail below)...can be modified with any of the below described approaches.    Bacterial strain with integrated defective...requires drug selection.  One drawback to this approach is that it requires the generation of bacteriophage...modification into a different background, if needed. This approach is best suited for modifying plasmids or BACs ...

... on genome sequencing. Used in concert, these approaches can provide the researcher with a reasonable ...(Fu et al. 2013; Pattanayak et al. 2013). This approach misses larger mismatches (e.g. six nucleotides... may be to integrate both biased and unbiased approaches. As suggested in a review by Tycko et al., 2016...a highly accurate manner. Using both of these approaches may not be necessary for every CRISPR experiment...Mark J., et al. "Evaluation of TCR gene editing achieved by TALENs, CRISPR/Cas9, and megaTAL nucleases....

...found yourself staring at a browser full of tabs, each open to a different antibody option. Or you may ... options in the Antibody Data Hub. Data reports  Each antibody submission is used to create a data report...Curating the data reports Just like our AAV Data Hub, each data submission we receive is checked by Addgene's... Rachel Leeson...