.... "Genome-wide recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library... will introduce you to this new technology, and guide you through the basics of CombiGEM-CRISPR experiments...with which one can design and synthesize CRISPR guide RNAs (gRNAs) for genome editing in large-scale has... has led to the rapid generation of genome-wide gRNA libraries that knock out (Doench et al., 2016; Hart...regulation in biological systems. The CombiGEM platform provides a means to create barcoded gRNA libraries that.... Ma, Hongming, et al. "A CRISPR-based screen identifies genes essential for West-Nile-virus-induced cell...Catch up on Your CRISPR Background with Our CRISPR Guide Pages Find CRISPR Plasmids Find More Tools for Combinatorial...

...Addgene’s plasmids are used with a wide variety of restriction enzyme-based cloning methods. Each method...traditional" restriction enzymes in that they cleave outside of their recognition sequence, creating four base...such as BsaI or BbsI), that are located on the outside of the cleavage site. As a result, these sites ...exploits the ability of Type IIS enzymes to cut outside their recognition site and permits DNA fragments...Gate cloning for inserting the appropriate oligonucleotides specifying a gRNA target sequence into a Cas9...amenable to "domestication", you might want to consider using an alternative method like Gateway cloning...cloning or Gibson assembly. Another important consideration is the design of flanking overhangs. Although...

...to use them, what they can tell you, and how to identify which one is right for you. Statistics – The ...permutation of pairs within the five is different. To identify significant pairings, additional tests, referred...hoc test, done after your initial analysis can provide information on specifically which groups are statistically...within your population? Sometimes this can’t be avoided, but if it can be, you can reduce the family-wise... p value less than or equal to the alpha value divided by the number of null hypotheses. Benjamini uses...uses a complementary approach which identifies a p value cut off. This method entails arranging all calculated... by finding the largest p value where its rank divided by the total number of comparisons performed multiplied...

...DNA bases without breaking the DNA backbone, sidestepping the problem of HDR vs. NHEJ entirely. The first...hypoxanthine (shown) is the nucleobase component of the nucleoside inosine. Created with BioRender.com. How ...al., 2016). Target-AID acts similarly but not identically to BE3, modifying a 3-5 base window 18 bases ...editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated with... created HF-BE3, a base editor containing high fidelity Cas9 variant HF-Cas9 (Rees et al., 2017). HF-BE3...positions 4-7. ABEs 6.3, 7.8, and 7.9 display slightly wider editing windows of position 4-9, although editing...Although the lab did not conduct comprehensive genome-wide studies of ABE specificity, their other experiments...

... in genetically identical mice. Thus, scientists must genotype each animal to identify the ones with the...-lox Read our Genome Engineering Guide Learn more in our CRIPSR guide ...traits and genetically engineered mouse models provide researchers with useful and adaptable tools to ...background, which means that each mouse is genetically identical and all mice of the strain are homozygous at nearly...inserted gene may have errors introduced to the nucleotide sequence during the DNA repair process. The addition...reporter gene, which replaces the targeted gene and provides a way to track successfully engineered cells. ...

... 2021,  with over 70,000 articles in PubMed, the idea that DNA methylation is linked to disease has now...tools to specifically edit methylation sites.  Step aside, transcriptionally active dioxygenase  When I started...Recently, however, a particular tool has gained widespread popularity – and notoriety – as a tool of choice...   Let me leave you with a few practical considerations: Design your gRNA to overlap the CpG of interest...interest. Having the CpG directly inside the gRNA sequence is a great guarantee that DNMT1 will be blocked...Cas9 Resources at Addgene.org Read our CRISPR Guide to Epigenetics References Charlton J, Jung EJ, ...Galonska C, Charlton J, Mattei AL et al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. ...

...You’ll want to set up your blot with a polyvinylidene difluoride (PVDF) membrane, which is sturdier and...Western blots are a great tool to identify a protein of interest in a complicated solution like cell ...already run your blot using nitrocellulose, you may decide you have nothing to lose by trying to strip and...abundance of your targets isn’t too different, consider antibody affinity: stronger binding antibodies...matter what stripping methods you throw at it. Ideally, dry your membrane once before any detection steps...(βME) 100–115 mM = 0.7%–0.8% (v/v) Breaks disulfide bonds HCl Adjust pH to 6.7–6.8 Adjust ...addgene.org Western Blot Protocol Western Blot Protocol Video ...

...Chemogenetics has become a popular tool in the field as it provides a noninvasive method for manipulating cell activity... fast-acting optogenetic tools. The advantages provided by chemogenetics are instead that it is noninvasive... crossing the blood-brain barrier.  Another consideration when picking ligands for chemogenetic studies...response you aim to manipulate. It is important to consider that the same receptor can elicit different responses...muscarinic receptor derived DREADDs has been clozapine-N-oxide (CNO). However, concerns have been raised over the...Pharmacokinetic and pharmacodynamic actions of clozapine-N-oxide, clozapine, and compound 21 in DREADD-based chemogenetics... Gomez J, Ramirez M, Hu X, Galvan A, Basu J, Michaelides M, Sternson S (2019) Ultrapotent chemogenetics...

...isotype choice. Target and application should be considered The first question you may have is whether a ...monoclonal primary antibody (A). To boost signal, consider using subclass specific secondary antibodies instead...strength (Fig. 1). Monoclonal primaries If you do decide on a monoclonal antibody you may wonder, “Will ...immunoprecipitations. Your sample species should also be considered when choosing an antibody. In some applications...options are available to you, it may be worth considering an antibody raised in a different species.    ...      Table 1: Isotype considerations when choosing antibodies   Complexity is a key factor...with a distinctly conjugated secondary antibody. Consider too that some secondary antibodies can cross react...

...sequence or a sequence provided by the depositor. Depositing labs often provide insert sequences or annotated...to check the quality of other services that we provide, including viral preps and pooled libraries. As...this changeover had no impact on our ability to provide requesting scientists high quality data! Addgene's...culture to prepare it for sequencing, we feel confident that these few minor mismatches usually don’t ...differs from what we would expect given the data provided by the depositing lab, we call these “quality ...Sequencing and Analysis Platform to Assess the Identity of Recombinant Adeno-Associated Viral Preparations...

..., loci were identified for 61 such diseases. Now single gene alleles have been identified for 4,163, over...surveillance of repetitive gene families and provide a way to guide inheritance that might be less error prone...plasmids, especially those of the CRISPR variety (see slides from my talk below). Largely organized by Dmitry...of items, only a small number of which I could identify. There were dozens of different mushroom varieties...how the NIH sees the future for genomics, and President Obama’s Precision Medicine Initiative and how ...shRNA silencing of one circular RNA they have identified resulted in a cell migration defect. They have...

...magnetoresponsive proteins. Motivated by curiosity Although a wide range of fluorescent proteins and optogenetic tools...possible only in non-biological conditions. So, they decided to search for a magnetic response among other proteins...Hayward et al. 2024 under CC-BY license.   Video 1: Fluorescence movie of E. coli expressing MagLOV...imaged area measures approximately 82 mm across. Video courtesy of Andrew York and Maria Ingaramo.   MagLOV...fluorescent proteins.  Currently, the team is working to identify how MagLOV’s structure changes under magnetic ...microscopy tools for biological applications across a wide range of topics. He embraces open science, self-... for?”, and while Andrew and Maria have lots of ideas and are happy to speculate, they know they can’t...

...with low fidelity polymerases and the cost of synthesizing longer oligos, which could be avoided using restriction...into E. coli for propagation. However, while not widely known, plasmid assembly can be achieved in vivo... of a plasmid, order primers that bind at either side of the undesired portion, designed to amplify the... Mutagenesis is performed by designing primers astride the old codon, amplifying the whole vector. The...homology, so performing more complex procedures can be aided by extending homologous sequences (to 25-30 bp)....colonies for simple cloning procedures. Use high fidelity DNA polymerase This will minimize random errors... blog posts Read our blog post on 5 factors to consider before choosing a cloning method Check out all...

...envelope have high infectivity in a wide range of cell types, so they’re ideal for work with primary cells or...envelope protein (Env), usually from VSV-G due to wide infectivity, is often used as a substitute for HIV..., used both for targeted modification and genome-wide screening.   Lentiviral vectors in the clinic Since... check out our lentiviral plasmid and biosafety guides before beginning your experiment. Let us know how...PMCID: PMC4273606. 4. Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem...Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional Resources...Resources on Addgene.org Check out Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors...

...plasmid from a bacterial culture, it is important to consider your experiment.  It will dictate the amount of...260 and 230 nm ratios should fall ~2 - 2.2, and provide an indication of purity versus chaotropic agents...agents as guanidine thiocyanate and guanidine hydrochloride used during plasmid extraction. Figure...be entirely endotoxin free. Endotoxin is a lipopolysaccharide of the cell wall of Gram negative bacteria... For these reasons commercially available kits provide extra washing steps to favor the removal of this...want to utilize your plasmid DNA, so that you can decide what will be the optimal type of plasmid preparation...Brush up on molecular biology protocols using our video collection ...

...restriction sites into which you can clone your 20 nucleotide guide sequence of choice (for generation of a gRNA... to the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables selection...plasmid post transfection. The 2A "self cleaving" peptide causes the ribosome to translate Cas9-2A and the...the Puromycin resistance gene as separate peptides. PX459 This is the vector backbone. This particular...cleavage site). Finally, in addition to the 2A peptide on its C-terminus, SpCas9 is also fused to a 3X...Resources on the Addgene Website Visit our CRISPR Guide Pages Browse Our CRISPR Collection Find gRNA Design...

... we set out to streamline this process. We take pride in facilitating the exchange of not just the physical...that allows you to conduct your experiments with confidence. Benefits of the UBMTA: Peace of mind Addgene...University Technology Managers (AUTM). This MTA provides many benefits for both the providing and receiving... lists below for more details. Benefits to the provider (organization / lab that owns the material) ...publish without editorial comment or review by the Provider Limited liability Addgene makes it easy Even...can get tricky. Here’s how Addgene helps: For Providers: One-time Deposit: Simply deposit with Addgene...the UBMTA and are happy to agree to it. Their confidence and familiarity with the UBMTA result in quick...

...recombinant Adeno-associated virus (AAV) has been widely used to deliver genetic tools to the brain because...neuronal circuits. Epitope tags are short antigenic peptide sequences, attached to a protein of interest (POI...simian virus 5-derived epitope (V5), the synthetic peptide FLAG, the synthetic streptavidin-binding strep-...both (Viswanathan, 2015). For Looger’s team, an ideal probe should combine the solubility, cell tolerance...imaging experiments, the second color is usually provided by red fluorescent proteins such as tdTomato or...smFPs can label fine neuronal structures with high fidelity and at lower concentrations than GFP. smFPs enable...N-cadherin in neurons and show that smFPs fusions provided better labels than fusions to 3 or more HA tags...

... live it out, but ideally without asking too many (annoying) questions. Identifying and solving problems...was aware that such roles existed, but did not consider them for myself. I started picturing myself in...Other Addgenies and Their Careers in Our Career Videos What's required for a role in technical support...any incoming requests or problems - there is considerable diversity in the types of questions customers...consuming and/or less technically challenging. Alongside there is a second-line team that takes on cases...problem. It may not always be straightforward to identify those crucial details because there are just so...Visit the Addgene Careers Page Check Out Our Career Videos ...

...When choosing beads, there are several factors to consider. Agarose beads are sponge-like and vary in shape...shape and structure. Their porous surface provides a large area for binding, but antibodies conjugated ...to the capture antibody and is immobilized. The ideal antibody concentration to use will vary between ...antibody is functioning as expected. In order to identify any non-specific interactions, perform parallel... is used to isolate the protein of interest. The wide availability and high specificity of anti-epitope...appealing, but the disadvantages must be carefully considered before using. For example, the tag may affect...routinely use the chromatin IP (ChIP) technique to identify DNA binding proteins involved in histone modification...

...advantages and disadvantages of each, and some considerations when using the method. ELISA basics All ELISAs...methods. For enzymatic signal detection, users provide a substrate that the enzyme converts into a detectable...those studying low abundance antigens may want to consider alternative detection methods.      Figure...conjugated primary antibody. The reporter reacts with a provided substrate and produces a signal that is directly...can be challenging since scientists must first identify two independent antibodies that work well together... of conjugated antibodies   Additional considerations No matter what ELISA method you choose, it ...sample of wild type cells, purified protein, or a peptide. The positive control will confirm that the procedure...

...though. Typically, you will identify a ‘scientific mentor’: someone who provides career advice and offers... and identify any gaps or questions you have, then bring them to your mentor. Pro tip! Consider physically... a quality mentor, and rightfully so! They can provide you with high-level career advice, open doors for...Training follow up After your training session (ideally in the next few days or weeks), look over your ...protocol, or maybe you would like to take pictures/videos of a machine set up while your mentor is demonstrating..., your bench mentor’s training time is time set aside just for you – so do your best to show up as your...

... will outline a simple method for selecting the guide RNA, validating its efficacy in vitro, and using...d like to manipulate in mind. Now it’s time to consider where you would like to attack this gene. Should... When the target sequence was digested by sgRNA guided Cas9 endonuclease, homology dependent repair (HR... It can take roughly 3 months from the day you decide to do a gene modification experiment in mice to...have your F0 generation (barring life’s little accidents and stuff ups). This is a huge improvement on ...production of mouse lines that could be used to study a wide variety of phenomena, it’s possible that we’ll learn...of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases." Genome research 24.1...

... in lower lentiviral titers. Another factor to consider is the age of your culture post thaw. In general...lower titer virus. In such cases you may want to consider concentrating your viral prep. Storage can also.... For long-term storage, viral preps should be divided into single-use aliquots, and stored at -80°C. ... start to a successful infection, one point to consider is how the virus is being titered. Most titering... that routinely produce lentivirus may want to consider testing a variety of cation and serum sources .... “Mycoplasma contamination of cell cultures: Incidence, sources, effects, detection, elimination, prevention...About Viral Vector Elements Learn How to Do Genome-Wide Screens with CRISPR Lentiviral Vectors Additional...

...and stability. TRX and signal peptide tags help proteins with disulfide bonds fold properly. If a TEV ...the subcellular localization and the number of disulfide bonds in your protein - two parameters that affect... and efficient multiplexing. pCri plasmids are divided into two sets, each of which includes plasmids ...designed for these systems may include a signal peptide (SP) that routes the protein through the secretory...secretory pathway to improve folding and disulfide bond formation. If one prefers expression in E. coli, the...thioredoxin A (TRX)-containing vector can improve disulfide bond formation. pCri also includes a special feature...

... Genomics Institute partners with companies to provide technological resources for those studying rare...be used to accelerate rare disease research and provide hope to patients suffering from these devastating...Schiavi plans on using C. elegans as a model to identify SMN1 interacting partners and perform whole animal...of mesothelioma sufferers. This disease is less widely understood and often misdiagnosed. Like other forms...mesothelioma tumor samples that they have used to identify candidate genes as drivers of peritoneal mesothelioma...Meothelioma Cancer Alliance Website.  We are proud to provide support to Drs Di Schiavi and Nabavi and will continue...., et al. "PP09. 1–2352: Systematic review of incidence and prevalence of spinal muscular atrophy (SMA...

...ultimately flattening the curve, it has become evident that widespread testing is crucial in controlling this ...PCR using Taq Polymerase and then transcribed to provide RNA substrates for Cas13. Cas13, upon finding complementarity...RPA, this technique uses Taq polymerase which is widely available and less expensive. Instead of using ..., field-deployable nucleobase detection and identification using FnCas9 (FELUDA) Sensitivity: – 110 femtomolar...FnCas9, this is also a good way to diagnose single nucleotide variations. Also, FnCas9 cleaves in a range of..., field-deployable nucleobase detection and identification using FnCas9. https://doi.org/10.1101/2020.04.07.028167....org Get the basics about CRISPR in our CRISPR guide Browse our entire COVID-19 plasmid collection ...

...Over 25 million people worldwide are currently infected with the lentivirus HIV-1. Today, HIV-1 can be...the virus doesn’t completely disappear; it just hides in latently infected cells. To truly cure HIV-1,...removal of the HIV-1 genome from infected cells, an idea that became much more realistic with the advent ...Cas9/gRNA expression. A pooled analysis found no evidence of Cas9 cleavage at predicted off-target sites...into a therapy, these papers present tantalizing evidence that an HIV-1 cure may be within our reach. Similar... disease that infects over 250 million people worldwide. As we’ve previously seen in the CRISPR field,... Learn to Use CRISPR Pooled Libraries for Genome-Wide Screens Additional Resources onf Addgene.org ...

...different ways to clone DNA fragments which have provided appealing alternatives to restriction enzyme cloning..., I highly recommend watching this entertaining video created by our friends on the Cambridge 2010 iGEM...fragments to incorporate such regions of homology. This video gives a helpful demonstration of how to use Snapgene...the process works best with fragments over 200 nucleotides. This may be because the exonuclease could chew... through an entire fragment shorter than 200 nucleotides before the annealing and polymerization steps...methods (Kahl 2013). However, Gibson assembly isn’t ideal for synthetic biology standards, which rely up on...Type IIS restriction endonucleases) to cleave DNA outside of the recognition sequence. This requires that...

...deletions are defined using 2 loxP variants (only identical loxP sites can mediate recombination.) The deletions...Optimizing the system: Brainbow-3 Although Brainbow provided neuroscientists with the vast array of colors ...variety of fluorescent proteins to find those with ideal characteristics (low aggregation, high photostability...produces a minimum of 8 colors. The use of AAV provides spatial and temporal control without the need ...Drosophila and zebrafish. Brainbow in Drosophila has aided in the mapping of neural circuits, such as connections... tool to mark and follow single cells due to the wide array and high stability of colors. Sanes and Lichtman...further refinements of the Brainbow technique will provide important insights into the complicated physical...

...open a heart to look at the inside. We do a lot of interactives and videos, as well, to supplement the...addition to testing those experiments. “We also do videos demonstrating an experiment or a technique to students...gel [for] electrophoresis or doing a dissection video. I did that in January. This involves going in front...the courses encompass what they want to teach.” Besides working with students and faculty, Caitlin also...would be to take any opportunity in the lab or outside the lab: write!” Caitlin did this by editing 15...on Facebook. “I think this helped me say with confidence when I interviewed, ‘I write science. I edit ...

...various conditions. The first thing you'll need to decide when running your experiment, is which vector will...will discuss some of the features you may want to consider as you make your choice. The first and most important...selection marker or antibiotic resistance. Next, consider the expression level of your construct: would ... lentiviral or a retroviral backbone. This is considered to be a more robust expression system and has...are several types of yeast expression vectors. Consider your applications and experimental design. You....1 - TRC cloning vector). Check out our complete guide to empty backbones for additional details and guidance...Plasmids 101 blog series Addgene's Molecular Cloning Guide & Other Plasmid Protocols List of common epitope...

...their interest in science more generally to people outside of academia. Here are my recommendations on some...magazines or newspapers or other places online, and consider how those stories are being told. Twitter can ...science blogs written by practicing scientists. 3)   Consider starting up your own twitter feed or talking about...Close the gap between science and society 6)   Consider participating in a future SciFund Challenge. While...motivated more by the desire to help close the widening gap between science and society by connecting ... Santa Cruz, has been turning scientists into confident and connected science writers since 1981. That... both your plasmids and your research stories. Consider practicing communicating your research by guest...

...interests. We've outlined some important factors to consider and summarize helpful advice from the successful...Achieve Success in Academia". Important factors to consider when choosing a research project After joining...and goals. The following questions can serve as a guide when selecting a research project. What will the... are available in order to realize your project ideas. What mentors are available for guidance, intelligent...fungal biology.” Connie Cepko advises students to consider their goal of PhD training when selecting a research... good judgement with the current technology to identify a problem, asking what approach will I take and...touching a pipetteman. Take the time to generate new ideas and think about them critically before diving in...

... nucleases from re-identifying it (3). Surprisingly, few genes have been identified that neutralize CRISPR...Addgene’s CRISPR Guide or find CRISPR/Cas plasmids for your research. Do you have a cool idea or application... process the CRISPR loci to generate RNA, which guide a Cas nuclease to the specified exogenous genetic...engineering. The diversity of the CRISPR/Cas systems provides powerful lines of defense against invading phages...technology? Let us know.  References Ishino Y, Hideo, S, Makino K, Mitsuko A, Nakata A. J Bacteriol. ...

...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ...structural understanding thereof have the potential to provide improvements to existing technologies based on ...PubMed PMID: 29915061.        A new method to identify proteins occupying a specific genomic locus: Caspex... as an affinity tag. This ‘Caspex’ construct is guided by a pre-defined sgRNA sequence to a specific genomic...preprint posted July 4, 2017 A new method to identify proteins occupying a specific genomic locus: C-BERST... and H2O2, APEX2 labels electron-rich amino acid side chains (e.g. Tyrosine) with biotin. These biotin-labeled...spectrometry allowing a researcher to specifically identify proteins that are enriched in the 20 nm radius...

...kinetics of the target ligand. Another important consideration is whether the affinity of an experimental protein-ligand...Proteolytic removal of the fusion protein could provide a more relevant kinetic analysis, but the trick...immobilization through biotin-avidin binding could provide a true measurement of high affinity (i.e. sub-nanomolar.... MBP has a similar function as GST in that it provides a method to express a foreign protein in bacteria...biotinylate the lysine residue in the AviTag. As a side note, we found that adding a TEV linker dramatically...that we couldn’t find an all-in-one plasmid that provided an easy way to biotinylate a fusion protein in...biotinylation and purification strategy outlined here provides a basis to determine binding kinetics using a ...

...transposon? The piggyBac™ transposon was originally identified in moths in 1983, but wasn’t successfully used... that have negative impacts on fitness. One consideration when making a transgenic cell line with the ...the piggyBac™ system, these challenges can be considerably reduced. The piggyBac™ transposon system can...donor-plasmid, you could use a single-stranded oligodeoxynucleotide (ssODN) as the donor [6]. The problem with... his specialized knowledge to help researchers identify the right technologies and strategies for their...sites is cytotoxic for proliferating cells and provides a simple tool for conditional cell ablation.,”...Zinc-finger nuclease-induced gene repair with oligodeoxynucleotides: wanted and unwanted target locus modifications...

...without changing them. Thus, it has been widely adapted for a wide variety of applications: base editing,...fusions localized to specific loci based on the guide RNA sequence. A year later, they developed CRISPRainbow...RFP) to the genomic loci of interest. Pairs of identical hairpins within one gRNA will result in a blue... chromosome-specific loci in the genome and the guides are relatively unstable.   Figure 1: CRISPRainbow...deliver CRISPR-Sirius into cells. They named this guide sgRNA-Sirius-8XMS2 which will recruit the HaloTag...Find plasmids for your research Read our CRISPR guide ...

...research from bench to bedside is, in addition to thorough ethical considerations, in making sure that ... our CRISPR guide Find CRISPR plasmids by function Check out our genome engineering guide ...scientists and the public together to build consensus guidelines, but there was never a specific timeline for ...shuffling our feet on necessary actions to review and decide on the future of genome editing technology. We ...Harley and Lexi, and a clutter ofoutdoors-only spiders, all of whom are named Charlotte, all of whom are...

...brilliant ideas on how to solve many therapeutic and pre-clinical problems! Who knows, perhaps the ideas spurred...students play, you can observe, answer questions, and guide them to productive strategies while noting any gaps...their knowledge. Practically speaking the format I decided on was to have two teams of 4 people face each ...with them. My impression was that they had clearer ideas about gene therapy in the following lessons. And...freely share your projects with us! This exchange of ideas will help improve life science and biomedical education...molecular biology educational resources Find protocol videos to teach basic lab techniques Plasmid Land: The...

...yoga classes. Get outdoors. Taking a walk or a bike ride is a great activity for both exercise and clearing... times of stress due to #COVIDー19. Receive and provide support while being safe. @GHP_HarvardChan — Shekhar... (individual development plan) from Science to identify skills to work on for the next steps in your career...networking events. Download Addgene’s Science Career guide to learn more about career paths and things you ... Everyone. Plan out new experiments carefully. Consider submitting a registered report, where peer review...is based on the methods, not the results. Also consider using this framework to design your future experiments...Technique. Co-work virtually. With co-working, the idea is not necessarily to chat, but to mimic your usual...

...’ll be confident in what to expect. No system, including MTAs, is perfect, but it does provide consistency...1995: the UBMTA. Because biological materials are widely varied with many properties, having well-understood...Instead, we facilitate the signing of MTAs by the Provider and Recipient (in Addgene parlance, the depositor...plasmids are distributed under the UBMTA, which is widely used across academia. It’s our process that differs...to note that it’s the depositing institute that decides the terms of distribution for their plasmids. ...Webpage, help center articles, and our MTA YouTube video for more information. Happy transferring!    Many...

... AAV (rAAV) vector (Figure 1). Everything else outside the ITRs gets left behind.    Figure 1: Recombinant....com. ITR structure  ITRs are symmetrical 145-nucleotide sequences that flank the ends of the single stranded...Figure 2). Additionally, the ITR contains a 4-nucleotide Rep binding element (RBE) that serves as a binding...sequences. GC tails are often located immediately outside of the ITRs in the plasmid backbone and do not ...since analysis of individual reads allows for identification of non-dominant ITR sequences.  Restriction...Capsids More resource on Addgene.org Addgene's AVV guide AAV Viral Preps References  Earley, L. F., Conatser...Alexander, I. E., & Logan, G. J. (2019). A User’s Guide to the Inverted Terminal Repeats of Adeno-Associated...

... took classes in biochemistry, microbiology, epidemiology, and bioanthropology. Studying human health ...doing in the lab and the interactions I was having outside of the “ivory tower.” I’d spend the afternoon lovingly...survived this long without downing any dihydrogen monoxide (H2O). Studying science communication as a PhD...quality of the study design or by their preconceived ideas about the controversial medical issues. Participants...beliefs even though all three texts were otherwise identical. Likewise, when participants read an abstract ...in a behaviour called the “biased evaluation of evidence,” in which people (conscious or unconsciously)..., looking for reasons to dismiss them. There’s evidence to show that people with greater levels of science...

...This post was contributed by Deborah Sweet, Vice President of Editorial at Cell Press. Almost everyone who...some point. Usually it comes up when a researcher decides on a new project and begins by trying to reproduce... various different versions of checklists and guidelines to help authors understand what they are expected...NIH) now look for data management plans and for evidence of sharing for grant renewal—but there is clearly...hiring, funding, and promotion processes, I’m confident that everyone’s enthusiasm for actually making...Sweet from Cell Press. Deborah Sweet is the Vice President of Editorial at Cell Press.  She trained originally...editorial work to focus on a broader role as Vice President. Her current responsibilities encompass overall...

...Services - includes videos on general resume writing, as well as additional videos with emphasis for graduate...Relations Director for Curiosity Collider (hyperlink https://curiositycollider.org/) , which she co-founded, ...applying for the position?   Here are a few things to consider when you're preparing your resume for a non-academic...you only have two pages, you need to have a good idea of what should be highlighted on your resume, and...postings and look at the jobs you are interested in. Consider listing the skills you already have, and the skills...

...Microscopy, University of Amsterdam. Before you decide which car you want to buy, it is worthwhile to ...tables with fluorescent protein properties and provides a quick way to compare fluorescent proteins, e.g... the protein of interest by a 2A self-cleaving peptide (Goedhart et al, 2010). An explanation of the assay...proteins (with the same reference protein) under identical conditions, a ranking of the practical brightness...practical brightness. Therefore, practical brightness provides a better picture of what can be expected in a ...theoretical brightness. Consequently, it makes sense to identify a couple of promising fluorescent proteins and...mimicking the future application. In figure 2 we provide an example of the comparison of the practical brightness...

...Viral Service. With their support we are able to provide faster access to a larger number of viral preps...Karpova’s lab from Janelia Research Campus have provided valuable insight into AAV production. Finally...advice and encouragement. Why virus? Why did we decide to take on this project? Addgene’s mission is to...plasmids in a ready-to-use viral format. Having identified this need in the community, we approached it ...control, generates  educational materials, and provides viral vector technical service (details about ...expertise in answering questions on virus and helping provide educational resources on this collection. With ... we could ship internationally and be ready to provide extensive logistics support for export and shipping...

...collection that grows on a daily basis, the lists below provide a snapshot of relevant resources and a good jumping...the gene you’re looking for on this list, please consider searching directly on our website. In-exhaustive... from a Set of Mitotic Chromosomes. Samwer M, Schneider MWG, Hoefler R, Schmalhorst PS, Jude JG, Zuber...DW Cell. 2017 Aug 24;170(5):956-972.e23 Screen identifies bromodomain protein ZMYND8 in chromatin recognition...receptor signaling. Zhao K, Wang W, Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR Cell. 1998 Nov 25. 95(5)...chromatin and the epigenome. If you have been considering some new techniques, the posts below may steer...