...miscommunication, a lack of leadership, and inappropriate tools or writing approaches can lead to frustration, delay...Weill Cornell Medicine. Discussing data with colleagues can be more difficult during the pandemic era...can contribute and sketch on the board without leaving Zoom. The annotate function can also make the screen-sharing...cultural backgrounds.” This work outlines the need for clear communication, fair credit through authorship, and...ways of working, reconnect as human beings and colleagues, eventually encouraging purposeful and creative...Medicine in NYC and author of the book “The Salmon Leap for PhDs: Swimming upstream a career from academia...

...through October 19. The BioBuilder Learning Lab The BioBuilder Learning Lab is a fully equipped, state-of-the-art...and teachers in the U.S. and beyond. “I want to learn more about Synthetic Biology. Can you tell me where...capitalize on the excitement found through experiential learning. “Students may forget the technical details they... BioBuilder experience With its intentionally malleable curriculum, BioBuilder takes cutting-edge research...a project-based curriculum, BioBuilder expands learning outcomes by emphasizing the importance of critical...production of fire-resistant housing insulation, and cleaning up salt runoff from winter icing.BioBuilderClub...

... we've heard over and over from our users: "Please, please improve your plasmid maps!" After thoughtful... here to see the new map in action). Read on to learn more about the improvements we’ve implemented and...extraneous ORFs that cluttered our old plasmid maps. Clearly Annotated Features, Enzymes, and Primers - The ...easier for you to share plasmid information with colleagues, or explain plasmid features to a lab member ...Once you’ve identified a plasmid that you want to learn more about, updates to our sequence analyzer will...hover over any feature, enzyme, primer, or ORF to learn more information about it. For example, if you hover...

... Sample6 has changed focus and names, swapped leadership, and grown to 25 members.  What is Sample6?...competing in business plan competitions. After much learning, the focus of the application was changed, but...best, wins. He explained that entrepreneurs must clearly communicate their idea at various levels of complexity...industry vary drastically, he reassures that one can learn it by reading a broad variety of materials and constantly...million. The biggest challenge during this time was learning to communicate effectively to non-scientific audiences...are essential to the biotech startup puzzle: Nucleation of the team. The team is key, and the initial...raise money and/or validate an idea or product? Please let us know in the comments section! And read on...

...often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase activities.    ...site for integration is TA dinucleotides and it leaves behind the CAG DNA footprint from its terminal ...terminal sequences at the excision site after cleavage by the transposase. It has a cargo capacity >100 kB, although... TTAA and unlike other transposons, it does not leave behind a DNA footprint sequence after excision. ...   Applications of transposons Now that you’ve learned about some of the popular transposon systems, let...DNA into the pronuclei of fertilized eggs, which leads to random incorporation of that sequence into the...ijms22105084 Additional resources on the Addgene blog Learn about the INTEGRATE system for bacterial genome ...

...macroautophagy of the cytosol leads to accumulation of Pho8Δ60 in the vacuole, where it is cleaved to generate an active... molecular weight shift of Pho8 from its uncleaved to cleaved form is the final read out of this assay...cytosol, to the higher pH of the vacuole. Read on to learn more about prior methods for studying autophagy ...the vacuole via the secretory pathway, and then cleaved to generate an active form of the protein. To monitor...lysosome, since both Atg8p and LC3 are degraded or released once an autophagosome fuses with the vacuole or...

...that delivering RNA-guided nucleases or TALENs together with AdV donor DNA leads to a vast majority of AdV-modified...chromosomal region. The CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various... and demonstrated by Manuel Gonçalves and his colleagues at Leiden University Medical Center are now available...DNA for accurate genome editing with engineered nucleases. Nat Methods 11:1051–1057 . https://doi.org/10.1038...Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse...delivery of CRISPR/Cas9. Journal of Controlled Release 298:128–141 . https://doi.org/10.1016/j.jconrel...

...the RNases on your hands can degrade RNA. Keep a clean work area, which may include spraying your bench...Alternatively, glassware can be baked at 180°C for at least 4 hours.   If your final RNA sample(s) are resuspended...cell lysis.  In the late 1970’s, Chirgwin and colleagues showed that a strong protein denaturant, guanidinium...mindful of your extractions- if you need really clean RNA, it’s important to make sure that when extracting...effect of ammonium sulfate on the activity of ribonuclease A. Biochimica et Biophysica Acta (BBA) - Enzymology...active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299. https://doi.org/... Leah Schwiesow...

...requisite for a functioning lab, but it is for a scaleable lab. When you need to track the location, quality...post we’ll highlight some of the lessons we’ve learned over the years with the hope that our insights ...containers and contents Right from the get-go be as clear as you can be (there will always be confusion about..., tightly coupling them to their contents might leave you sad and confused when a tricky procedure failed...worm that requires a change of tube, there’s no clean way to separate the contents from the container ...etc.There are also going to be situations where you learn biology terms (plasmid, 5-prime, CRISPR, growth ...krisshamloo.com     Additional Resources on the Addgene Blog Learn about How We Use Barcodes in the Lab Get Tips on...

...the Addgene Blog Learn How Scientists Are Trying to Shape the Future of Research Learn How Notions of Scientific...studies with concrete examples of how gender analysis leads to discovery and innovation. One such case study... of their cells when doing experiments—which can lead to failed research. For example, an international...develop represent the whole human population. To learn a little more about where differences in effects...There is much work to be done! Researchers need to learn sophisticated methods of sex and gender analysis...Blogger Londa Schiebinger. Londa Schiebinger is a leading international expert on gender in science and technology...

...Ask openly for clear information on costs. Can you suggest some coaches? Yes I can. Please note that Addgene...to change jobs? Grow your professional presence? Learn a key new job skill? Finish your thesis? Knowing...that will help you. A good coach usually will be a clear communicator and make direct suggestions for improvement...She is also a very popular speaker and workshop leader. Lauren Celano of Propel Careers is not a professional...engagements and has many areas of specialty from leadership development to mentoring for women and under-...Guide Your PhD with An Individual Development Plan Learn a Little about The Academic Labor Environment from...

... 2019). Inhibiting DNA cleavage Example: AcrIIC1 binds to the HNH endonuclease domain of Cas9 and prevents...prevents target DNA cleavage (Harrington et al., 2017). Figure 1: Anti-CRISPR proteins (Acr's) can...loading, inhibiting DNA binding, or inhibiting DNA cleavage.   Five ways to anti-CRISPR proteins in your...timing experiments, the Doudna lab found that at least 50% of on-target Cas9 edits happens within the first...effects and cytotoxicity that result from excessive nuclease activity in tissues (Li et al., 2018). 4. Selection...Blog First time CRISPR user? Check out this post. Learn how to Choose the Best Cas9 Variant for Your Next...

...anymore. Say hello to PAM-free and PAM-flexible nucleases! In this blog, we will review Cas enzymes, which... uses a PAM with no sequence constraints, or at least has looser or different requirements. Luckily, there...Nonetheless, in an AT-rich region, these Cas9s may still leave you choosing between efficiency and location.   ...display higher off-target effects than SpCas9, a pleasant surprise since the PAM flexibility could in theory...activity of SpRY was validated in double stranded cleavage and base editing applications, with high editing... NG Cas9 NG - SpG Cas9  NG  - SpRY nuclease  C to T Base Editor SpRY A to G Base Editor SpRY...  Nishimasu, H., et al.,Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science 361,1259...

...implanted into a female animal for gestation. Nucleases that induce double-stranded DNA breaks, such as...cells, which are then injected into a blastocyst. Nucleases can also be used to engineer knockout mice. ZFN...non-homologous repair at the site of DNA damage. These nucleases are microinjected into a zygote, which is then...introduce point mutations. While using targeted nucleases to generate an engineered mouse model has advantages...targeting vector alone, some challenges still exist. Nucleases may cause off-target effects on regions of the... random process, zygotes edited using the same nuclease may not result in genetically identical mice. ...about Cre-lox Read our Genome Engineering Guide Learn more in our CRIPSR guide ...

...mEosFP exposure to UV light causes an irreversible cleavage near the chromophore, resulting in emission of...to blue light also inactivates the chromophore, leading to a non-fluorescent state. Dronpa can be reversibly...quickly inactivated and less photons are emitted leading to dimmer fluorescence. In fact, many of the early...are being emitted, making it easier to capture a clear image above background. The majority of PA-FPs come...disrupting cellular function and localization, or leading to protein aggregation. Another important parameter...light used to activate the first PA-FP does not photobleach the second, and vice versa. Finally, as every...contrast, PA-FPs offer a reduced potential for photobleaching and phototoxicity that standard FPs might induce...

...including mitosis, mitochondrial fission/fusion, nuclear import, and neuronal trafficking. Although FPs ...Overexpression of fluorescently tagged proteins can lead to improper protein localization, protein aggregation...power of this system lies in the ability of the endonuclease Cas9 to create a DNA double stranded break (...the tag, selectable marker, and connector to be released (Fig. 1, Step 2-3). Since donor plasmids of the...directed repair after a CRISPR/Cas9 induced break. Learn more about these constructs and the cell lines they...Cas systems used for endogenous protein tagging, please visit our website CRISPR/Cas Plasmids - Protein...PMC4889673. Additional Resources on the Addgene Blog Learn about Fluorescent Protein Timers Check out Photoactivatable...

...DNA could lead to transgene expression in recombinase-negative animals, producing misleading results.  ...the plasmid used to create the viral prep. In a clean viral prep, like AAV-44362 shown below, more than...from these known impurities, as was the case for clean sample AAV-44362. In addition to these expected ...number of hits to the same sequence (gene). In a clean sample, there are typically fewer than 10 hits to...receive the best AAV vectors for your research. Please use the comments section below to let us know if...Topic Page Subscribe to Viral Vectors Blog Posts Learn How We Use NGS for Plasmid Quality Control Resources...

... Developed by David Liu and colleagues, FokI-dCas9 expresses Fok1 nuclease domain fused to catalytically... to this system by engineering cells that lack release factor one (RF-1; Bacterial strain EcAR7) and minimizing...control for your experiments. For more information, please see Addgene’s information page for the Rinehart...neuronal signaling. For more information on DREADDs, please see the Roth lab's DREADD users blog. New ... expression plasmid that was used by Chen and colleagues for genome editing inXenopus tropicalis. (Guo...information on using the Platinum Gate TALEN Kit, please see the Yamamoto lab’s protocol for TALEN construction...mGRASP A Nature 2012 article by Jinny Kim and colleagues describes their efforts to map the location and...

...to see the dirty laundry exposed here, but it’s unclear how much they will actually care. The section of...technology. Though these case studies are “old,” they lead Zimmer to one of his most critical observations:...detection had over 1,000 authors. The chapter on deep learning addresses the dilemma of creating a technology...discussed The State of Science with the five Addgene colleagues who read it with me, the consistent response ...non-scientist alike was that the book left them wanting to learn more. There are individual paragraphs that may have...task to set oneself. As a scientist who enjoys learning a little bit about many different things (part...about a long time. For the scientists who like to learn everything about one specific topic, The State of...

... clinical gene therapy applications. Read on to learn more about the current (and future) applications... co-transfection of these plasmids, 293T cells released transgene-containing lentiviral particles into...and they showed that expression persisted for at least thirty days. Although adeno-associated viral vectors...activation from the 5’ LTR) and Rev (facilitates nuclear export of transcripts). A different viral envelope...safer, this system may be more cumbersome to use and lead to lower viral titers due to higher plasmid number...Addgene Blog Your Lentiviral Plasmid FAQs Answered Learn How Lentiviral CRISPR Libraries Enable Genome-Scale... on Addgene.org Check out Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors for...