...SciComm by doing SciComm Writing had always been a strength of Susan’s, but she didn’t know if she liked science... of a team.” It took her two years to find her current position at Fred Hutch because her skill set wasn...connections she made through this program lead to her current job. “One of my colleagues in the class was like...Hutch magazine. Getting pulled in all of these different directions is sometimes at odds with the space...

... vector team is publishing research, hosting conferences, and are expanding our catalog of materials to...appearance and functionality are happening and this strengthens our image in the world. Addgene is now as much...practices and benefits and these are making a difference. We serve scientists around the globe and our...Thanks to all the Addgenies and congratulations to current and alumni Addgenies on our success. Thanks to ...

...extra care when performing RNA extraction. RNA is inherently not as stable as DNA- it is single stranded and...ultracentrifuge.  Figure 1: An outline of the different steps in RNA extraction. Researchers at the... Reagent®, RNAzol®, QIAzol®  and sold by many different companies). TRIzol® is an all-in-one acid-guanidinium-phenol... ensure DNA contamination is not an issue.  References                 Allewell NM, Sama A (1974) The...

...antibodies Conventional antibodies are a non-renewable reagent - Their production requires animal sacrifice...produce - Hybridomas offer an alternative and renewable source of monoclonal antibodies, but they are ... their use. Each RANbody is detectable by two different tags: a reporter and an HA tag. The reporter comes... link to its sequence in the comments below. References Sanes, J.R., & Yamagata, M. (2018). Reporter-...

...but where the kinases are located, their signal strength, and how long their signals last all impact transduction... or “on.” Dronpa’s color thus indicates MEK1’s current state (Green when off, dark when on). Figure 1...function of the protein and pdDronpa are preserved. References 1. Zhou, Xin X., Linlin Z. Fan, Pengpeng Li, ...28232577. Figures 2-4 were modified from this reference. 2. Regot, Sergi, Jacob J. Hughey, Bryce T. Bajar...

...mammalian systems has been in question. Three different labs have recently published work that provides... gene activation  and base editing, whereas a different set of mutations (the “V3.1” version) works best...miniature Cas system that utilizes a Cas12f from a different species, Acidibacillus sulfuroxidans, AsCas12f1...of up to 10kb but it is also capable of coding concurrent small insertions to allow in-frame deletions ...

...sixty seconds of silence–but it was thorough and coherent.  To be honest, if my paper hadn’t been ridiculously...to stop talking until I could present something coherent or to quickly string together all the words that...screeching feedback. You’re muted on Zoom. Your video conferencing software freezes and then crashes. You’re muted... a clear, informative presentation, speak at conferences, or even just provide quick updates during a ...

...if you need a very pure sample. Because binding strength is proportional to the length of the tag, with...Antibodies.) Unfortunately, for some proteins there just aren’t any good antibodies available. In these cases, ...antibody posts from our blog!  Resources and References  Resources on addgene.org Addgene Antibody Guide...ChIP Antibodies 101: Introduction to Antibodies References Hochuli E, Bannwarth W, Döbeli H, Gentz R and...

...experiment moving. Happy editing! References and Resources References Hendel, A., Bak, R., Clark, J. et...talk about their precious RNA samples. RNA is inherently less stable than most macromolecules used in ...region of the gRNA. These gRNAs can be cleaved and rendered useless after less than a minute of exposure to...

...the target strand and the gRNA. Created with BioRender.com. To decrease the electropositivity of the...mutations). Although the precise mutations were different than those found in eSpCas9, the effect on specificity...and/or therapeutic settings. Will combining the different mutations found in each of these great tools further...continued improvements in genome engineering! References 1. Slaymaker, Ian M., et al. "Rationally engineered...

...scientific manuscript will require numerous other references to scientific literature to substantiate the ...literature related to the topic. Finding reliable references and sorting out what they mean is no small task...focus in on a particular conclusion. Many papers aren’t always written this way and there is a greater ...inspiration and accountability. There are a few different ways to automate the process as well. Sign up ...

...without permanently altering the DNA, CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) are...study ncRNA, CRISPR systems offer a number of different options. By carefully considering your ncRNA of... you more about its function.  Resources and references Resources on addgene.org Addgene's CRISPR Guide... CRISPR enzymes for transcriptome engineering References Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna...

...CRISPR experiment. Happy cloning! References and Resources References McGee, A. V., Liu, Y. V., Griffith...chosen because of their broad applicability in different model systems. What’s the error rate? The Doench...Table 1 has a list of Fragmid destination vectors currently available. Table 1: Fragmid destination vectors...

... one. As a starting point, I would recommend conferences like Lab Robotic Interest Group (LRIG). There...customizability. It has a large deck that can house many different types of devices. As long as a device fits on ... Software for Biology Tips from the Repository Trenches: Using Barcodes to Track Samples Resources at...video library Check out our molecular biology reference pages ...

... we haven’t even imagined yet. References and Resources References Andrews NP, Boeckman JX, Manning ...consist of a mixture of antibodies recognizing many different epitopes. (Middle) Monoclonal antibodies produced... Trimmer JS, Görnemann J, Glanville J, Wolf P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P...

... recombinase/photodimer pairs. Created with BioRender.com In their recent ACS Synthetic Biology publication...fold more photostable and as bright to mVenus, the parent YFP, and is the most photostable YFP to date. As...and reduced RNA-off target footprint compared to current N-terminal linked SaCas9 ABE variants. It's also...specific tight affinity glutamate sensor from the Loren Looger Lab now available in AAV1. Get GFAP.SF-iGluSnFR.A184S-AAV1...

...options as well as the benefits and drawbacks of different methods (for comprehensive protocols for all of...used in the transfection; if preparing several different lentiviruses in parallel, this approach should...that standard conditions be used when comparing different batches of virus. In addition, some viral preps...step of a successful transduction experiment. References 1. Kutner, Robert H., Xian-Yang Zhang, and Jakob...

...you can make an informed choice! References and Resources References Bondeson, D.P., Mullin-Bernstein,...are titratable. With these tags, protein levels aren’t limited to ‘on’ and ‘off’; varied levels of drug...profile their effectiveness with over a dozen different proteins. In the end, you can’t predict with absolute...

...chromoprotein and how is it different from fluorescent proteins? Before going into the difference between chromoproteins...research Read our Molecular Biology Reference Find plasmids for FRET References Blanquet RS, Phelan MA (1987...

...Also, keep all liquid containers closed when they aren’t in immediate use. This will prevent spills, but...could change their growth rates and morphology, rendering their continued use inadvisable for experiments...in a lab where multiple people are working on different things, this will help you avoid cross-contamination... Immunology. You can find Sana on LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling...

...species, like S. aureus and N. meningitidis require different PAM sites than SpCas9, in this case, NNGRRT and...single PAM recognition. Created with BioRender.com. After 24 days of phage propagation, Hu ...percentages, with fold changes compared to SpCas9 in parentheses. The development of xCas9 3.6 and 3.7 shows ...is working for you in the comments section! References Hu, Johnny H., et al. "Evolved Cas9 variants ...

... Let it incubate Check the bacterial strain Differences between E. coli host strains can impact plasmid...might want to include an extra wash step or use a different strain. For most applications, Addgene uses and...inadequate plasmid DNA yields. Why would volume make a difference? In the simplest of terms, more volume = more...conditions play a critical role in plasmid DNA yield. Different plasmids and strains will vary in their optimal...

... ever found yourself bamboozled by all of the different kinds of information on our plasmid pages? Well...depositing lab, although you may have an entirely different use in mind! This section of our plasmid pages... you to use in your Materials and Methods and References sections when citing the use of your colleague...e.g. if it was derived from a plasmid made in a different lab - or one obtained from Addgene!).  It also...

...anymore. The kit contains sequences for several different fluorescent proteins. Visualizing fluorescence...use our promoters and terminators, but then they aren’t sure how to convert their own coding sequence to...with the Athena SWAN Charter and it has really strengthened the network of female scientists. Twitter has...for those who go to work in cancer biology.  References: A Golden Gate Modular Cloning Toolbox for Plants...

...known quantities of DNA. Additionally, an AAV reference, a virus with a known titer, can be used to confirm...both results would be valid. This is why the AAV reference is critical. It allows you to determine if your... PCR (ddPCR) does not require a standard or a reference, meaning saving on reagents (and time)! AAV titering...becoming a popular option for PCR experiments.  References Furuta-Hanawa, Birei, Teruhide Yamaguchi, and...

...alleles, or introduce a premature stop codon to render an RNA nonfunctional. In their quest to build a...are present in the gRNA spacer, as ADAR will preferentially edit an adenine if the template has a cytosine...(Abudayyeh et al., 2019). To avoid some of the inherent drawbacks for RNA editing of natural cytidine ... Valley contributed to updating this article. References Cox, David B.T., et al. “RNA editing with CRISPR-Cas13...

... adaptations of this system have been made in different fields of study. In addition to the identification...purification with an anti-Cas9 antibody. These different tag systems and affinity purification schemes ...dystrophy (FSHD), a disease for which there is currently no cure (Campbell et al., 2018). Find plasmids...please also see our interview with Dr. Fujii. References Campbell, Amy E., et al. "NuRD and CAF-1-mediated...

...dimerized linear ssRNA strands (which can have a different genomic sequence from one another) — by various... with lentiviral replication.  Resources and References More resources on the Addgene blog Viral Vectors...popular lentiviral plasmids γ-Retrovirus Guide References Cullen, B. R. (1992). Mechanism of action of ...Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives. Viruses, 13(7), 1288. https...

... applications. Read on to learn more about the current (and future) applications of lentiviral vectors...facilitates nuclear export of transcripts). A different viral envelope protein (Env), usually from VSV-G...plasmid number. For more information on the key differences between the 2nd and 3rd generation packaging ...use lentivirus in the comments section below! References 1. In vivo gene delivery and stable transduction...

...when separated (2), and that DBD and TAs from different systems could be combined effectively (3). In ...a more vigourous screen is to use a number of different bait and prey vectors. This has been shown to ...split ubiquitin system has been devised (8). References 1. Casadaban M.J., Martinez-Arias A., Shapira...Science. 1986; 231:699-704. PubMed PMID: 3080805. 3. Brent, R., and Ptashne, M. A eukaryotic transcriptional...