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...for scientists working at home, and you will still find this useful. Now that we’ve been doing this for ...safe space. No matter why, now is a great time to find a few colleagues to work together on career and ... “the ability to assess and initiate things independently” and “the power or opportunity to act or take...format is good for everyone including those that find events stressful. Video chatting makes it pretty...effectively is a valuable skill. There are many different kinds of grants that you can apply for. If you are in ..., your PI is likely working on a number of them. Find one to apply to or offer to help with lab grant ...Opportunities Open Education Database: 100 Places to Find Funding For Your Research University of Florida:...

...still invested in seeing how they do it.    Finding Conflict in Science  In all science communication...conflict into your work, but one of my favorite is to find it within the science itself - within that process...trying to understand your completely unexpected findings, or understanding why your experiment has not ...not worked…again…for the seventh time….   I find that once I sit down and talk to a scientist about their...paper was actually from a completely unexpected finding in a different experiment, this tool was developed...fodder for science communication. The trick is to find one or two roadblocks that sparked creativity, changed... a story around them. A good rule of thumb is to find the pieces that make you think, “Oh, that’s interesting...

... autophagic activity, and are a more reliable indicator of autophagic activity. The aim of this post is...emits red and green fluorescence. Following the induction of bulk autophagy, Rosella is targeted to the ...interest to track the autophagy of that protein. Find the cytosol-targeted and mitochondria-targeted Rosella...nm to 440 nm serves as a measure of autophagy induction: when Keima is in the cytosol, this ratio is low... this biosensor can’t be used with fixed cells. Find the mitochondria-targeted Keima variant at Addgene... in yellow puncta but a decrease in red puncta. Find the mRFP-GFP-LC3 plasmid at Addgene! GFP-LC3-RFP-LC3...fluorescence suggests a decrease in autophagic flux.   Find the GFP-LC3-RFP-LC3∆G plasmid at Addgene! References...

...student hasn’t been asked the question: “Academia or industry?” Once academia was the clear answer with a well-defined...web-based, career-planning platform called the Individual Development Plan (IDP) and uses it to match qualified... qualified scientists to jobs in industry, academia, and government. This great resource allows the undecided...and other professions increases your chances of finding a well-suited job. EAs shouldn't be viewed as an...interests: Use career planning tools such as the Individual Development Plan to generate a list of potential... more information Follow people in academia, industry, and government on social media (Twitter, Facebook...potential job Join LinkedIn and other job sites to find open positions and view their required qualifications...

...targeting and sequencing millions of cells. For every individual sgRNA, you recover a minimum of 1,000 targeted... 5% off-target frequency is something to keep in mind, but likely will not have a significant impact on... many predictive online tools which can not only find gRNAs for you, but also score their probability ...probability of off-target events: CRISPOR, Cas-OFFinder, CCTop, and many more. While such software typically only...at mismatched guide sites, but not necessarily binding to these sites. Thus, these enzyme swaps will only...need to do your due diligence to ensure they are indeed a non-issue. Above we mentioned how to predict ...tested with some success. The major con is that binding by Cas does not always correlate with catalytic...

...expected fragments for that plasmid. This phenomenon indicates the presence of a multimer.   Fig. 1:...multimerization has been observed to occur in many different kinds of plasmids, taking many forms (Levene, 2009; Higgins... al., 2010; Cook et al., 2020). CGE uses laser-induced fluorescence (CGE-LIF) for detection and can provide...multimerization. One study by Bacolla et al. (2011) indicated that strain HB101 produced the highest yield of... at reducing multimerization and define recA-independent mechanisms. Avoid overgrowing the bacterial culture...-B DNA-forming sequences and WRN deficiency independently increase the frequency of base substitution ...26104344. Fishel RA, James AA, Kolodner R. recA-independent general genetic recombination of plasmids. Nature...

...seamless manner, leaving no sequences or mutations behind. Furthermore, piggyBac™ offers a large cargo-carrying...cells lines, including stem cells, without leaving behind any accessory DNA sequences [2], [3]. Simply put...integration [4], the donor cassette is removed leaving behind no accessory transposon sequences.  The use of ... plasmid after excision by PBx, thereby leaving behind only those cells with a Footprint-Free™ genome ...this method can be effective, a loxP site is left behind in the genome following donor removal, which alters...somewhere between one hundred and one thousand clones to find a cell line with the edit of interest. With the ...Cathomen, and K. Schwarz, “Zinc-finger nuclease-induced gene repair with oligodeoxynucleotides: wanted ...

...represent the fraction of that total with the indicated function. One huge reason CRISPR has become such... left the arduous task of virus production to individual labs, but now we’re very excited to provide ready-to-use...an entire lentiviral production protocol only to find that the low titer of her prep makes it unuseable...target these widely studied signalling pathways. Find Validated gRNAs to use with your Cas9 Virus Pooled...uses nearly 80,000 gRNAs to target around 20,000 individual genes. We provide enough lentivirus for you to... lentivirus production protocol here and you can find more tips and protocols for working with lentivirus...Viral Service Page Check Out Our CRISPR Guide Pages Find Viral Vectors ...

... are interested in understanding the mechanisms behind Mycobacterium tuberculosis resistance to pyrazinamide... and recombination-mediated genetic engineering (find more information on bacterial engineering using ...Devarshi Gajjar, The M. S. University of Baroda, India  Devarshi Gajjar has 10 years experience in the ...expressing fluorescent proteins will be used to tag individual strains of bacteria. Their interactions and growth...University of Ibadan, Nigeriaunder supervision of Dr. Akindele Oluwatosin Adeyi, University of Ibadan, Nigeria...PhD Candidate working under the supervision of Akindele Oluwatosin Adeyi at the University of Ibadan. ...regulation and diabetes will be expressed from plasmids (find more information on bacterial expression systems...

...found yourself frustrated reading through a paper to find that there is insufficient information about which...also designed to be persistent and exist online indefinitely. PIDs historically referred to objects that ...identify a plasmid available through Addgene and find verified information about each plasmid item. Given...associated plasmid page on the Addgene website and find specific information about the plasmid: name, description...and more. The plasmid page also helps researchers find other publications that have used the same plasmid... Addgene ID # and the information necessary to indicate that this ID is managed by Addgene. How to cite...down to the bottom of this plasmid’s page you will find that the “How to cite this plasmid” section now ...

...size of the insert. You can search NCBI for YGOI to find the particular reference sequence if necessary. ...The plasmid was digested with 2 unique enzymes (HindIII and BamHI) and run on an agarose gel. The resulting... (*). When the plasmid is digested with either HindIII and BamHI alone (lanes 4-5), there is a single ...size of the plasmid. The double digest with both HindIII and BamHI (lane 3) produces bands at 6kb and 1.2kb...bromide (EtBr) to your gel before pouring it. EtBr binds to the DNA and allows you to visualize the DNA under...vist Addgene's resource for plasmid verification to find additional tips and detailed protocols on topics...Resources: Addgene's Molecular Biology Reference Find More Protocols and Videos      ...

...Addgene.org Read our CRISPR guide Find CRISPR plasmids for your research Find resources and software for your...contains Cas9. The Madison lab also created an inducible SRIRACCHA system (iSRIRACCHA) where Cas9 expression...a 2-4 fold increase in the number of endogenous indel edits when compared to unsorted cells (Wen et al...leaving you with just your desired genomic edit.  Find SRIRACCHA plasmids at Addgene GEMCherry2, an improved...create a knock-out in human embryonic stem cells.  Find GEmCherry2 reporter plasmids at Addgene Overall ...Addgene blog Browse all of our CRISPR blog posts Find more about fluorescent proteins Get a refresher ...

...plasmids to help academic and industrial labs accelerate research and to help find a faster solution to the...methods Resources on Addgene.org Find all plasmids from Ginkgo Bioworks  Find COVID-19 plasmids for your research...current global pandemic in which scientists race to find vaccines, therapeutics, and other treatments to ...continually added as they complete QC and you can find all available plasmids listed on Ginkgo’s collection...global pandemic. And the reach of these plasmids is indeed global. Helping scientists speed up research is...

...Resources on Addgene.org Find base editors for your research Read our CRISPR guide Find plasmids for your research...modifying enzyme to the target site, and the deaminase induces the DNA base change without a DNA double-strand...for base editors is that the editing domain must induce the base change before the Cas domain lets go of...Cas9 domain to “hold on” long enough to the DNA to induce a base change. Because of this, ABEs are only weakly... DNA and off-target RNA editing rate of ABE7.10. Find the ABE8e plasmids at Addgene! Why engineer and ...adenine base editors can correct all four of these kinds of point mutations, in theory converting the disease-associated...

...novice and indicate why other neuroscience newbies might want to get into the field. Find Optogenetics...neuroscience but are afraid you don’t have the right kind of biological expertise to get into the field, there...require a bit of expertise and can be difficult for individual labs to make efficiently. For select viral vectors... to get some ideas for future experiments and to find tools that will make your transition into the field... If there’s a plasmid or AAV tool that you can’t find, you can always suggest a plasmid to us, request...Resources at Addgene.org Addgene's Viral Service Find Retrograde AAV Browse Optogenetics Plasmids ...

...won't cut it for bioinformatics. All kinds of biologists are finding themselves using computational approaches...Additional Resources on Addgene.org Find Protocols for Your Next Experiment Find Our List of CRISPR gRNA Design...data sets (myself included) and we are faced with finding the best system to document these types of analyses...bioinformatics. In fact, there are endless ways and everyone finds their own. Here, I simply outline what works for...not apply or may not work for you. The best way to find a system that does is through trial and error, and...

...mentors as one key to their success. But how do you find these elusive teachers, supporters and advisors?...trainees more about peer mentoring. They want to find mentors to help them develop their skills and to...accountable for their development plans. However, they find it awkward to ask someone for the time commitment...mentors available to go around. My advice has been to find 5 interested peers and form your own peer mentoring... steps to forming your own peer mentoring group Find 5-8 colleagues that you think might be interested...diversity of perspective. Sometimes it is helpful to find peers that share a similar direction or problem....potluck dinner. Invite guests from a lab, career or industry of interest – learn about their career paths and...

...aiming to increase an antibody’s binding specificity, you can perform binding assays against the target antigen...Moffitt Cancer Center. Engineering usually calls to mind building things, lots of math, and maybe heavy machinery... and antibody fragments. Fab: fragment antigen binding; Fc: fragment crystallizable; IgG: immunoglobin... antibody, but don’t have a specific mutation in mind. Generating a random library of mutants using error-prone...your mutant library and increase the chances of finding beneficial mutations. In other words, structure-guided...conditions, such as pH or temperature, can be adjusted to find the best-performing mutant antibody in those conditions...direct screening via flow cytometry for improved binding under defined environmental conditions. These methods...

... Resources on Addgene.org Find ready to use AAV vectors at Addgene. Find AAV plasmids to use in your... other. When Cre is introduced to the system it binds one pair of lox sites, let's say the loxp pair, ...expression?   Figure 1.  Cre-recombinase binds to either pair of lox sites, let's say the lox2272... the level of spontaneous recombination. This indicates that the amount of leaky expression can largely...recombination levels as high as 1-3%. In light of this finding, we now include guidance on the plasmid page of...levels of TH mRNA without expressing TH protein indicating that there is promoter activity. Additionally...Route for Adeno-associated Virus (AAV) Entry Independent of AAV Receptor. J Virol 92. https://doi.org/...

...typical part-time or 9-to-5 job. I had to ask to find out what a “postdoc” is, and that “PI” stood for...experimental design, and execution. Can lead to an independent project. Lab technician / Research assistant...next career stage (ex: grad school, med school, industry) Other roles you might see Lab manager: ...be unsure, it helps to have a vague sense of what kind of research you would be excited about. I know quite... in high school, but I’ve learned the value in finding mentors wherever I go. This story by Dr. Erika ... to Become a Scientist While Poor” Keep an open mind as you explore opportunities The world of science...doing scientific research. It’s okay to change your mind about what exactly you want to do—I started college...

...similarly useful across the diverse kinds of proteins that one finds in cells. High yield of reaction ...freedom to test out lots of combinations, to end up finding something surprisingly useful. To apply these principles...HaloTag modules, which can connect proteins to any kind of small molecule probes with different functionalities...Exchangeable HaloTag Ligands HaloTag Reversible binding of fluorescent ligands to HaloTag PCV-Cas9, Cas9.... Applications are quickly expanding with these kinds of Click Biology reactions. For basic research, ...proteins. Split inteins are particularly useful for a kind of cellular computation, making possible multiple...

... Resources on Addgene.org Find Empty Backbones with Fluorescent Tags Find Plasmids for Subcellular Localization...are visible. These whorls can be quantified for individual cells, and the number of 'normal' looking cells...the results of the OSER assay, you should keep in mind that the presence of whorls is probably concentration...monomeric behaviour in cells (Shaner et al, 2013; Bindels et al, 2017). Still, the data presented in papers...is detected by Constantini et al (2015) and by Bindels et al (2017) but not by Cranfill et al (2016))....build a valuable resource with tools and data that indicate the performance of (a set of) fluorescent proteins...

...days prior to the implementation date, our data indicated that, while 69% of all postdocs were expected ...stakeholders. However, while most institutions are indeed going ahead with their planned raises, some are...had that increase cancelled. Rounding out the remainder, 2% of postdocs who weren’t going to get a salary... reversed their position and decided to allow individual departments to go ahead with the raises as they...detrimental effect on the morale of postdocs, and indeed we have been contacted by a number of postdocs ...to reach out to institutions with a statement indicating that we encourage them to keep raising salaries...Variety of Science Careers Resources on Addgene.org Find Career Opportunities at Addgene  ...

...Background with Our CRISPR Guide Pages Find CRISPR Plasmids Find More Tools for Combinatorial Engineering... Wong. The complexity of biological systems can hinder our attempts to study and engineer them, but what...Gilbert et al., 2014; Konermann et al., 2014) individual genes for studies interrogating their functions...incredibly useful for the efficient generation of individual (and multiple) genetic perturbations in single...sequences via oligo synthesis using the format indicated below and pool-clone them into the pAWp28 storage...identifies genes essential for West-Nile-virus-induced cell death." Cell reports 12.4 (2015): 673-683....

... alternative evidence and discard inconvenient findings. Yet, we cannot prosper in a world where our medicines...ominous political climate in which American scientists find themselves today, and where Canadian scientists ...the public conversation and to tell the stories behind our research.  That's why I marched for science...University of Michigan campus speakers thoughtfully reminded us to use the same critical lens on our scientific... Aguilar Galileo wrote, “now you would not hide behind these two fingers the ninety-nine cubits of Aristotle...promote harmful policies that benefit polluting industries.In March, three climate scientists and an economist...specific, technical manner, so much so that we often find it difficult to convey the excitement and importance...

...a look in a typical lab freezer, you’ll probably find lots of different plasmids containing the same protein...Leu28Cys mutation in eDHFR does lead to covalent binding of the TMP-tag ligand (Gallagher et al., 2009)....based on a bacterial haloalkane dehalogenase that binds a synthetic chloralkane (CA) ligand. The current...before investing in one approach or the other.   Finding the right tag for you Protein-based tags typically...Promega (though if you are comfortable with it or can find a colleague to help you, synthesizing your own ligands... experiment, and explore Addgene's collection to find self-labeling fusion proteins or cloning backbones...An In Vivo Covalent TMP-Tag Based on Proximity-Induced Reactivity. ACS Chemical Biology, 4(7), 547–556...

...dividing cells. Find tips on using gamma-retroviral vectors in our Gamma-retrovirus guide and find popular packaging...tetracycline inducible, 3rd generation lentiviral vector backbone from the Shih lab. You can find more lentiviral... vectors. These vectors can induce combinatorial constitutive or inducible gene overexpression, knockdown...-lox system is a technology that can be used to induce site-specific recombination events and is widely...

...of upfront research you have to do to find said robot and to find out how you can get one. As a starting...at Addgene so we’ve been putting some work into finding robots that’ll take a little labor off our hands... pod can pick up 8 tips and position the tips indepently making it easy to spread out your samples. This...live bacteria. For these shipments, we’ve needed to find a way to purify DNA from many bacterial samples ...same time as opposed to preparing each sample individually. Enter the BIOROBOT! This robot is called the...Barcodes to Track Samples Resources at Addgene.org Find protocols for standard molecular biology techniques...

... on Addgene.org Find CRISPR Plasmids for Your Research Interested in light-inducible Cas9 activity? Check..., 2018). For example, AcrIIA4 mentioned above indicates that it’s the fourth Acr protein discovered that...CRISPR activity one of three ways: Inhibit DNA binding Example:  AcrIIA4 blocks Cas9’s interaction with...2019). Inhibiting DNA cleavage Example: AcrIIC1 binds to the HNH endonuclease domain of Cas9 and prevents...such as inhibiting crRNA loading, inhibiting DNA binding, or inhibiting DNA cleavage.   Five ways to ...of CRISPR activity Regulating Acr proteins by inducible promoters, light, or small molecules could achieve...

...interest  Figure 1. When ABEs are in action they bind DNA so that the TadA domain can edit the short single-stranded...creation of accidental insertions and deletions (indel) by the catalytically impaired D10A nickase from... efficiency in a human cell line while reducing indel frequency more than 90% compared to ABE7.10. Choosing...changes in cellular DNA and RNA dependent or independent of the gRNA used (Gaudelli et al., 2017). The...in this work did not cause genome-wide, guide-independent off-target deamination, an important attribute...0491-6 Additional resources on the Addgene blog Find more blog posts about base editing Learn about another...Addgene.org Get the basics of CRISPR in our CRISPR guide Find all plasmids for base editing in our collection ...

...vector blog posts Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all...do this, the group developed a new strategy for finding retinal cell-type specific promoters. Most existing... mouse cell type specific transcription factor binding sites interspaced with random sequences.   Strategy...transcription start site. TFBS: transcription factor binding site. Image from Jüttner et al., 2019 with permission...that was active in the outer part of the retina.  Find the AAV synthetic promoter plasmids! Comparing synthetic...Read more about using AAV for retinal gene therapy Find more about precise neuronal targeting using AAV ...

...trials involve identifying a therapeutic gene, finding a means to deliver it and identifying a suitable...significant clinical efficacy against a form of blindness known as Leber’s congenital amurosis (LCA). This...RPE65. It is characterized by childhood onset of blindness and was thought to be untreatable until 2008. ...their efficacy:  Pre-existing immunity: Many individuals already have wt AAV in their bodies and may therefore...interfere with virus-cell interactions and seriously hinder gene therapy outcomes. Anti-AAV antibodies developed...decline in vision with gene therapy in childhood blindness." New England Journal of Medicine 372.20 (2015...Quality Control Process Resources on Addgene.org Find ready-to-use AAV in Addgene's Viral Service Check...

...this protein is well suited for genome editing.  Find Cpf1 Plasmids at Addgene   Image attribution...red) or LbCpf1 (blue). Mismatch positions are indicated below each graph. Figure adapted from Kleinstiver... and LbCpf1 ribonucleoproteins (RNPs) failed to induce off-target editing in a cell culture model. What...Nat Biotechnol. 2016 Jun 6. PubMed PMID: 27272384 Find plasmids from this publication at Addgene. 2. Kleinstiver...Nature Biotechnology (2016). PubMed PMID: 27347757. Find plasmids from this publication at Addgene. 3. Zetsche....1016/j.cell.2015.09.038. PubMed PMID: 26422227 Find plasmids from this publication at Addgene. 4. Makarova... Guide Pages Check Out MpAgo from the Doudna Lab Find Validated gRNAs ...

...flexible linker is a methyllysine binding domain (chromodomain), which binds selectively to lysine-methylated...like the potential devastation of the palm oil industry through epigenetic effects on the Cover of Nature...maternal copy of the gene is expressed in normal individuals (in a parent-of-origin-specific manner), but ... the histone-derived peptide, the methyllysine binding domain causes a structural change (facilitated ...682-688. PubMed PMID:12404098.  4. Lin, Chi-Wang, Cindy Y. Jao, and Alice Y. Ting. "Genetically encoded ...documentary PBS Additional Resources at Addgene Find the Ting Lab plasmids at Addgene Browse More Fluorescent...

... is the only right way, and Indecisives, who keep changing their minds and have trouble making decisions...categories. In addition, I also learned about two other kinds of difficult supervisors: Hands-off super busy ...work is just one of the hundred things on their minds. When you come to a meeting with a clear agenda,...poor mentors (super-agreeable friendly types, indecisives, and super busy types). If you supervisor ignores...you are advancing career, you will become the independent, assertive, and proactive person that all employers...a Good Mentor Resources on the Addgene Wesbite Find plasmids to Expedite Your Experiments and Achieve...

... pet gives you a clue to help you find the next picture. If you find the next correct photo, you will ...1 below!) You will find five photos this way. At the very last photo, you will find a link to a form where...our recently redesigned blog. 🐕 Your task is to find them, scavenger hunt style. 🔎 Here’s how the scavenger...scavenger hunt works works: You will be trying to find pictures of Addgene pets with Addgene-branded clothing...

...In order to bind DNA, Cas9 and other CRISPR enzymes require a short PAM sequence adjacent to the targeted... researchers have engineered alternative Cas9s binding distinct PAM sequences. Now, Hu et al., working...create point mutations, also has a strict editing window about 15 nt upstream of the PAM. Even if there’...ω-dCas9 could recognize a library PAM, it would induce transcription of gene III, permitting phage propagation...xCas9 activity with wt SpCas9 in mammalian cells, finding xCas9 3.7 displayed about the same editing rate...loci displayed a broad range of PAM sequences, indicating they may be less predictable than SpCas9 off-...Resources on Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids for Mammalian Systems Learn about...

...Additional resources on addgene.org Find Optogenetics Plasmids Find Biosensor Plasmids Visit the Fluorescent...multiple other labs have reproduced their EGFP findings since the initial report in 2023.)     ...changes in response to a ~25 mT magnet. Gray regions indicate times when the magnetic field is on. Image reproduced..., in response to a ~10 mT magnet. Gray regions indicate times when the magnetic field is on. Image reproduced...enhanced for future MagLOV-based tools, just as light-induced conformational changes are the basis of many optogenetic...collaborating, the York Lab is eager to hear from you. Find the MagLOV plasmid here!   References and resources...

... RNA (gRNA) sequence. Users can design gRNAs to induce the break at a specific genomic site and, using...screen, it is expensive and time consuming to individually generate gRNAs and homology arm repair templates...promote homology directed repair after a CRISPR/Cas9 induced break. Learn more about these constructs and the...for tagging endogenous proteins with an auxin-inducible degron (AID) tag to generate proteins which can...repressors, can be included in the repair template for inducible alteration of protein expression levels. CRISPR...Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron Tagging with Short Homology Donors.” Cell...Resources on Addgene.org Check out the SapTrap Kit Page Find Other Fluorescent Protein Kits Browse the Fluorescent...

.... Click on the gene names in the figure below to find plasmids containing that particular gene. ...) and Mark Prescott (Monash University). You can find more plasmids for labeling your subcellular structure...signals not visible to the human eye and are indifferent to subjective color intensity perception. Open...colocalization relies upon the detection of 2 independent fluorescent signals and their potential overlap...muli-color imaging. Furthermore, one must bear in mind that the resolution of light microscopy is limited...interest in a healthy or diseased cell. We hope you find the resources discussed in this article useful for...21209361. PubMed Central PMCID: PMC3074624. 5. Bindels, Daphne S., et al. "mScarlet: a bright monomeric...