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  1. The Scientific Conference Poster Session: Tips for Success

    Type
    Blog Post
    Published
    Aug. 29, 2019, 12:50 p.m.
    ...most prepared of us can find ourselves talking to our posters, so if you find yourself in this situation...session, or it may be held at an unfamiliar venue. Finding your poster board well ahead of time ensures you...at the actual venue. Introduce yourself: I always find it odd when I walk up to someone’s poster, and then... a lap around the posters and read the titles to find the ones you’d like to revisit. Strategically traverse...give the “elevator pitch,” of his/her project – kindly let the presenter know you are very interested ...either seek out the presenter after the session or find him/her on social media or through their affiliations...who presented on a topic related to your research? Find a way to get in contact with the presenter after...

  2. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ... weak gRNAs (CFD < 20) bind almost not at all, and strong gRNAs (CFD > 80) bind almost as much as perfect... when designing gRNAs Find other CRISPR software Resources on Addgene.org Find CRISPR plasmids and resources...rest of this article, we'll explain the thinking behind the tool, its key features, how it works, and how...we began using it internally. Guiding principles behind CrispyCrunch While CrispyCrunch may evolve over... not available publicly. Furthermore, we did not find any tools which were optimized for HDR, except for...typically advised to mutate the PAM to which the guide binds. However, if the PAM resides in a coding region,...single base pair may not be enough to prevent re-binding. Lastly, the sequence inserted by HDR may inadvertently...

  3. Of Myc and Men

    Type
    Blog Post
    Published
    Jan. 19, 2023, 2:15 p.m.
    ...specific epitope that your antibody binds. For example, Myc1-9E10 binds EQKLISEEDL (pronounced “equi-kli-...Resources on Addgene.org  Find recombinant anti-c-Myc [9E10] at Addgene Find plasmids with the myc-tag...same antibody developed almost 40 years ago? Well…kinda. In some cases, antibody providers are distributing...to protein engineering. As a result, you can now find Myc1-9E10 in a variety of isotypes like rat IgG1...this trusty tool doesn’t end there: you can also find anti-Myc [9E10] Fab fragments and single-chain variable...accessibility and the variety of formats you can find it in likely mean it will remain an important tool...tool for the foreseeable future. These newer findings simply add to our understanding of how this antibody...

  4. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...survey. When asked if they could detect indels (an indication of gene disruption ability) with NgAgo, ...surveyed here could not achieve indels with the NgAgo system indicates that it is very tricky to use and...many researchers were having difficulty achieving indel formation (gene disruption) with this new technology..., one respondent out of 88 confirmed successful indel formation at a target site. Also, 41 respondents...dead in the water. On the bright side, successful indel generation and epitope knockin was achieved by a... a few respondents suggesting that NgAgo can indeed be optimized for genome editing in mammalian cells...transfections with the ssDNA guide, although many may find this inconvenient. Another possible reason for the...

  5. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...interference, can reversibly exert their effect independent of gene copy number. RNA interference (RNAi) ... 1). As a result, gene silencing can be easily induced by exogenous application of long or short dsRNA...applications. Figure 1: Schematic of an RNAi-inducing transgene. Typically, the desired target of RNAi...experiments that require exogenous protein, successful induction of RNAi only requires the presence of dsRNA. In...phenotype. Long hairpin RNA The easiest approach to induce RNAi involves the expression of long hairpin RNA.... Each construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene...more specific than long hpRNAs, they must be individually engineered and empirically evaluated for their...

  6. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...the transposase enzyme binds the transposon’s long terminal repeats (LTRs), induces a double-stranded break...A DNA footprint is left behind. When the transposon-transposase complex finds its target site, it integrates...surveys the genome and binds its target DNA sequence. The transposase complex binds the mini-transposon,...for integration is TA dinucleotides and it leaves behind the CAG DNA footprint from its terminal sequences...and unlike other transposons, it does not leave behind a DNA footprint sequence after excision. piggyBac...Transposons are also less likely than viral vectors to induce an immune response and are also easier and cheaper...

  7. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...maintenance involved. Also it was really fun to find individual colonies growing out in the 96-well dish. My...transduced with Cas9 expressing lentivirus at the indicated multiplicity of infection (MOI). Anti-vinculin... the initial stable cell pool (pool) and from individual cell expansions (monoclonal, 1-5). Wild-type ...control GFP gRNA (BRDN0000561167) (80034-LV) were individually used to transduced stable pools of Cas9-A549.... I used existing BRAF gRNA viruses, but you can find a list of validated gRNA plasmids on our website...about (which could affect the ability of the gRNA to bind). Verifying genomic edits with the surveyor assay...the 428 bp Surveyor nuclease cleavage product indicates a gene edit. GFP gRNA-transduced Cas9-A549 cells...

  8. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    Published
    Aug. 30, 2022, 1:15 p.m.
    ...Antibodies Resources on Addgene.org Find recombinant antibodies at Addgene Find plasmids encoding recombinant...targets, preventing your primary antibody from binding (Fig.1). The level of cross-linking that occurs...temperature, so you may be able to recover good antibody binding by tweaking those parameters, but in some cases...So, while organic solvents may improve antibody binding for some targets, they can destroy it for others... 1: Choice of fixation method impacts antibody binding. Formaldehyde cross-links the protein (red lines...results by not permeabilizing as you’ll prevent binding to any target protein that is still on the inside...reveal otherwise hidden epitopes. However, keep in mind that you will still contend with many of the same...

  9. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    Published
    Aug. 20, 2015, 2:30 p.m.
    ...through winning and losing funding, and by finding our individual niches within the field of grants. What...want to be an independent grant consultant, building your area of expertise will help you find work and target...serve time and resources. 3. Get a mentor, find your people Find someone who is working or has worked as ...career as a Grant Professional has been a long and winding path for many in the field. The vast majority of...completed. To enter and advance the profession, you can find professional development, certification, degrees...career. Do you have a passion for the arts? Do you find satisfaction in helping your local food shelf raise...experience and to build upon your existing skills.  2. Find your niche Identify and develop an area of expertise...

  10. Increasing Your Work's Visibility with Addgene: Citation, Search, and Collections

    Type
    Blog Post
    Published
    March 10, 2016, 3:30 p.m.
    ...their work (H-index for instance), but these rely upon other researchers being able to find their publications...that another researcher finds useful, that researcher will be able to easily find the fruits of your labor...depositors, Addgene strives to make it easier for you to find the plasmids and publications relevant to your research...only does citing plasmids from Addgene help others find them later if they need to use them in their own...similar projects and should make it easier for them to find your work later. We also hope that this feature ...instance, on our Optogenetics Collection page, you can find tables containing plasmids for expressing different...Collections Pages Suggest Plasmids for the Repository Find Plasmids for Your Research  ...

  11. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...-throughput sequencing to identify genome-wide binding patterns of the target protein. Both IP/MS and...bottleneck, however, to these IP-based assays is to find (or generate in-house) a pre-validated antibody ...tag-specific antibody (Jarvik and Telmer 1998). Indeed, this method can be used at high throughput because... transcription factors in a short (3-week) time window; this was possible because reagents could be purchased...Ensembl genome browser: https://www.ensembl.org/index.html Design crRNA/ssODN: Using Ensembl Transcript ...recent article by Clarke et al shows that crRNAs binding to the template strand (strand used by RNA polymerase...knock-in efficiency CRISPR reagents had crRNAs that bind to the template strand. Therefore, I suggest that...

  12. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    Published
    March 31, 2020, 1:15 p.m.
    ...  Antisense RNAs encoded on the plasmid either indirectly inhibit replication through inhibiting translation...machinery proteins, or directly inhibit replication by binding to the origin of replication and blocking replication...the first, the replication initiation factor RepA binds to the iteron to initiation replication. However...However, in iteron-containing plasmids,  RepA can also bind to its own promoter and inhibit its own transcription...handcuff” model, iterons of two separate plasmids bind to the same RepA molecules, forming a “handcuff”...region on the plasmid, a centromere-like region binding protein (the CBP), and a partition NTPase (Schumacher...Schumacher, 2012).  During cell division, CBPs bind to the centromere-like region on each plasmid and pair...

  13. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    Published
    Feb. 17, 2022, 2 p.m.
    ...Development and Discovery As part of their quest to find, characterize, and develop new CRISPR systems and...tests samples of new bacteria from anywhere they can find, whether the samples are from a rain gutter or the...lab for testing, they weren’t sure what they would find… but it was safe to say the sample had their attention...inhibition when the system was retargeted. This finding was surprising – in the most common RNA targeting...further experiments strongly supported their initial findings.  The team originally suspected that this new ... any gene editing treatment would have to be individually customized to the patient's specific mutations... via Site-specific Targeting Elements (PASTE). Finding the right integrase for this new approach proved...

  14. Transferable Skills Guide: Creativity

    Type
    Blog Post
    Published
    Jan. 18, 2022, 2:15 p.m.
    ...   Thinking Outside the Box: Individual Approaches Whether it’s finding the source of contamination or...innovation. If you identify as a non-creative or if you find yourself in a creative rut, read on!   Fig... a colleague. To innovate, you’ll have to focus. Find yourself a block of time and a quiet space where...When they were instructed to use the three-ring binder as a starting point, the ideas generated were hardly...get around the lab faster that way— but the idea behind the study holds. To generate novel ideas, start...Team Approaches At some point or another, you’ll find yourself needing to brainstorm with a team. One ...t had time to process on their own. You may also find your meetings run smoother when people have thought...

  15. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows indicate natural cellular...the endogenous enzyme is not efficient enough to hinder adenine base editing. Once base transversion editors...C-terminal domain of Cas9, which narrowed the editing window and reduced A → G bystander edits. Their final ...these adenine transversion editors produce a lot of indels (up to 40% at one site tested with AYBEv3!). Although...substrate for a base excision repair pathway. The team behind AYBEv3 introduced the first deaminase-free base...transversion editors, this editor had relatively high indel rates ranging from 4.7% to 30% at tested sites. ...deaminase-free C → G editor with a different editing window than existing CGBEs. This distinct approach means...

  16. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ...distributors (Table 1), in China, Korea, Japan, Brazil, India, and Mexico. Table 2: Addgene distributors Distributor... Mexico e-nnovation Life Sciences 2020 India Sinapse Biotecnologia 2022 Brazil   ... large percentage of these shipments arrived in India at room temperature. Partnering with our distributor... items to countries with a distributor. Arrows indicate the year we entered into an agreement with a distributor...accessibility to our repository by making it easier to find materials and learn about them. As early as 2007...packaging systems, and luciferase expression. On individual material pages, we added curation fields for ...Collections Lentiviral Plasmids Tetracyline Inducible Plasmids Empty Backbones Bacterial Expression ...

  17. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...antitoxin) where CcdB inferes with DNA replication by binding the GyrA subunit of DNA gyrase and trapping it ...in DNA breakage and cell death. Conversely, CcdA binds to and blocks the activity of CcdB. Applications...of protein synthesis. The antitoxin MazE, which binds and inhibits MazF, is continually cleaved by the...control of the AOX1 promoter whose expression is induced by methanol. Once linearized, the delivery vector... where recombination has not occured, methanol induces the production of the toxic MazF leading to cell...triggering cell death. However, the sok antisense-RNA binds a sequence downstream of the mok SD resulting in... as an orally administered diagnostic that can indicate the presence of liver metastasis by producing ...

  18. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    Published
    June 27, 2018, 1:16 p.m.
    ...chromatin structure at your locus of choice. Find the Tet-inducible caspex plasmid here. Myers SA et al. Nature...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...technologies based on polymerase processivity. Please find the MarathonRT plasmid here! Zhao C, et al. RNA.... methods. Since the anti-GFP nanobody is able to bind fluorescent proteins derived from A. victoria the..., YFP, mCerulean, or other related fluorophores. Find all of the nanobody toolkit plasmids here. Buser... the authors specifically investigated protein binding at the TERT gene, but this method enables the unbiased...fused to APEX2 is expressed from a Tet/Shield1 inducible CMV promoter and targeted to a sequence of interest...

  19. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    Published
    March 26, 2019, 1:40 p.m.
    ...Learn about Addgene's viral service Find viral protocols and videos Find AAVs for your research ...intensities that change in response to calcium binding. There are a few versions of these jGCaMP7 sensors...small neuronal processes like dendrites and axons. Find the 7b variant of jGCaMP7 in AAV1 at Addgene! See...but exhibits larger fluorescent responses to GABA. Find the iGABASnFR and its F102G variant in AAV1 at Addgene...KGC-GFP-ER2]) of this membrane-embedding voltage detector. Find Archon1 and its cre-dependent version at Addgene... encoding a fluorescent-tagged molecular tool.   Find pAAV-FLEX-tdTomato, pAAV-GFAP104-mCherry, pAAV-mDlx-NLS-mRuby2...Also coming in our new 20 ul trial size! Voltage indicators Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST...

  20. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

    Type
    Blog Post
    Published
    Sept. 21, 2023, 1:15 p.m.
    ...problem! But to be sure that your secondary is not binding endogenous IgGs, you should include no primary ...antibody controls in your experiments. If you do find high levels of background staining by the secondary... Strategies for avoiding background secondary binding Direct immunohistochemistry (Figure 2A)  How do...do you avoid having your secondary antibody bind endogenous IgGs? Don’t use one. If it is possible, using...antibodies as normal. Ideally, your secondary will only bind to your primary in this scenario because the endogenous...species and 2) these fragments will be more likely to bind (and block) the same epitopes recognized by the ...sample so the secondary antibodies aren’t free to bind endogenous IgGs. (C) A third solution is to use ...

  21. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ... over the world, and the question on everyone’s minds is: could NgAgo replace CRISPR? Such a drastic shift...proteins play an important role in RNA interference, binding RNA guides to cleave foreign RNA. Argonaute proteins...homology-directed repair when a template is supplied. Find Plasmids from the NgAgo Publication at Addgene Cas9...observe differences in cleavage efficiency. If this finding is replicated in larger studies, it will represent... If you’re looking to disrupt gene function via indel formation, this property could be an advantage. ...fidelity would prevent other small nuclear DNAs from binding NgAgo, it might make it difficult to use NgAgo ...Biotechnol. Epub 2016 May 2. PubMed PMID: 27136078. Find plasmids from this publication at Addgene. 2. Swarts...

  22. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    Published
    April 9, 2019, 1:13 p.m.
    ...known as steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding sites or motifs...POI). To indicate the sequence, people often use terminology like “C-terminal fusion,” but I find this very...increases avidity because it doubles the number of binding sites per unit.  There are several ways to measure...applications, many were concerned with steric hindrance by the fluorescent protein. Thus, scientist began...Felipe O., et al. "Spontaneous Cdc42 polarization independent of GDI-mediated extraction and actin-based trafficking... PubMed Central PMCID: PMC6381139. Costantini, Lindsey M., et al. "Assessing the tendency of fluorescent...et al. "Expanded dynamic range of fluorescent indicators for Ca2+ by circularly permuted yellow fluorescent...

  23. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ...These RNA duplexes bind to the RNA-induced silencing complex (RISC). After binding to RISC, the RNA duplex...strand usually being degraded. The target strand binds to the mRNA, which is then cleaved by RISC and degraded...Argonaute protein. This Argonaute protein is what binds the small RNA and either cleaves the target mRNA...sequence is highlighted in green, with red boxes indicating mature siRNA. Hairpin loop is in between the ...sequence is highlighted in green, with red boxes indicating mature miRNA. Hairpin loop is in between the ...stable knockdown cells lines, and you can usually find validated shRNA sequences for your intended target...RISC: Formation, action, and degradation of RNA-induced silencing complex. Molecular Cell, 82(1), 30–43...

  24. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    Published
    Dec. 2, 2019, 2 p.m.
    ...interest. Any proteins that bind the RNA motif will be biotinylated.  Find more details about RapID! EXCELL...biotin. Biotin, is a natural coenzyme, that strongly binds the glycoprotein, avidin or a similar protein, streptavidin...biotin molecules to dissociate from the ligase and bind to exposed lysines on neighboring proteins. Thus...organoid or in vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the development ...biotin ligase from A. aeolicus, which lacks the DNA-binding domain (233 amino acids) found in the E. coli version...fusion proteins to specific locations in the cell.   Find BioID2 plasmids here APEX2: A more sensitive ascorbate...Lam et al. 2014). They named this mutant APEX2.  Find the APEX2 plasmids! TurboID and minturbo: Rapid ...

  25. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...dystrophin reading frame. gRNAs bind in intronic regions, so low-frequency indels will not cause frameshifts...Check out Our CRISPR Guide Pages Find CRISPR Plasmids for Your Research Find Validated gRNAs for Your Research...concept for future preclinical and clinical work. Find Plasmids for Disease Research on Our Special Collections... function. Without functioning dystrophin, an individual experiences progressive muscle wasting leading..., but at least partially functional protein. Individuals with these mutations are often asymptomatic, .... With regard to off-target effects, each group finds very low to no off-target activity at the ten highest-ranking...cause obvious germline modification, an important finding given the controversy associated with such editing...

  26. 25 Work From Home Tips for Scientists

    Type
    Blog Post
    Published
    March 19, 2020, 1:15 p.m.
    ...science on. Practice self-care and stay connected Find time for exercise. Many studios and gyms are now... to support your favorite local exercise spot or find something new. We usually have yoga class twice ...great activity for both exercise and clearing your mind. And because the outdoors is so huge, these are ... help you prepare for sleep. At Addgene, our mindfulness group is sticking with their Headspace practice...Work on personal development Take the myIDP (individual development plan) from Science to identify skills...and set up activities that kids can do more independently. For example, they could do something like puzzling... kids. Plan out meals and snacks ahead of time.  Find learning activities for kids. For example, Skype...

  27. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    Published
    June 4, 2019, 1:02 p.m.
    ...proteins that bind serotonin. Unger redesigned the binding pocket of the periplasmic binding protein so ...are able to quickly sequence an individual’s genome. But when they find genetic variants, what does that...Get the basics on GFP Find microbiology blog posts Resources on Addgene.org Find fluorescent protein ...” Unger began with the a bacterial periplasmic binding protein, one similar to the protein which gave ...bioreporter such that fluorescent proteins expression is induced prior to inoculation with IPTG. In the absence ...

  28. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    Published
    April 7, 2022, 1:15 p.m.
    ...periplasmic glutamate binding protein known as GltI. When the resulting fusion protein binds to glutamate, conformational...at Addgene.org Find Fluorescent Protein Biosensors Browse All Fluorescent Proteins Find AAV Tools References...SF-Venus-iGluSnFR). iGluSnFR3: Improved Glutamate Indicators for Synaptic Imaging However, the first two generations...anchoring domains The original iGluSnFR in the pMinDisplay backbone is fused to a specific peptide segment...transmembrane domain in the mammalian expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol...pRSET: Bacterial expression vector, use with IPTG induction to express soluble protein CAG: Generic strong... 30377363 Aggarwal, Abhi, et al. . “Glutamate indicators with improved activation kinetics and localiation...

  29. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    Published
    July 27, 2017, 1:11 p.m.
    ...Beyond these common applications, researchers are finding new ways to apply fluorescent proteins everyday...award opportunities. Xin Zhou: Fluorescent Light-inducible Proteins (FLiPs) Fluorescent proteins can be applied...these photoswitchable enzymes Fluorescent Light-inducible Proteins, or FLiPs.  Xin later optimized these...spatio-temporally controlled as a key advantage over other inducible enzyme technologies. She hopes that other researchers...create a wide variety of enzymes and proteins with inducible activity and recently presented her work at the...sfGFP fused to a His-tagged ABC transporter known to bind to the HMO of interest. Fatima specifically proposes...with bacteria engineered to produce HMOs, sensor binding results in an increase in fluorescence of cells...

  30. 5 Great Apps for Lab Life

    Type
    Blog Post
    Published
    Sept. 22, 2016, 2:30 p.m.
    ...buffers to use and that indicates if a sequential digest is needed. The enzyme finder is also a very helpful...learning proper cell culture technique, I tried to find an app that could help me with calculations. This...to skim the alphabet on the side of the screen to find the sequence for your enzyme of interest. Lastly...with research! It is always important to know what kind of reagents you work with in experiments. Hence,...app has a search function that allows the user to find an MSDS for a lab chemical of choice. Now, I can...quizzed randomly about a hazardous chemical and can’t find your lab’s MSDS book, this is a great resource to..., J, X, or Z. Some alternative, similar apps: Findings Lab Notebook Cell Culture Split Calculator by ...

  31. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...If you’re cloning a plasmid, you’ll need a way to find the needle in the haystack: the one perfect clone...selection strategy is not enough on its own to help you find your desired plasmid. In many cases, you will need...survive and you will need to sort through them to find the desired clone. Do your selection, then screen...𝛽-galactosidase, it turns into an insoluble blue pigment. Find plasmids for blue-white screening at Addgene! Some...proper positive and negative controls for your PCR.  Find the details and learn more about colony PCR in this...into your insert. Check out our Vector Database to find plasmid maps of the more commonly used backbones...you can use to confirm your clones. You can also find a list of popular sequencing primers in our Molecular...

  32. 7 Tips to Secure a STEAM Internship This Summer

    Type
    Blog Post
    Published
    Feb. 2, 2021, 2:15 p.m.
    ...cannot offer. Furthermore, companies across most industries tend to hire their former interns to fill entry-level... in.     STEAMid is an app that helps you find internships and scholarships in different subject...to submission. When in doubt, consult the web to find the tools that can help you craft a solid personal...Letters of recommendation wisdom  Reach out to 2-3 individuals in your professional circle to vouch for the ... superb and detailed portrayal of yourself. Be mindful that January and February represent peak season...for internship applications. Thus these noble individuals are inundated with requests. The easier you make...        5. Application submission  Set a calendar reminder for every deadline and shoot to submit your application...

  33. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    ...functions. Photoswitches are engineered molecules that induce conformational changes with light stimulation to...activation have been developed to selectively study individual types of GPCRs with greater spatial and temporal...of the receptor, similar to what the endogenous binding partner would do. This allows precise control over...molecules (i.e. photoswitches) attached near the binding pocket of an engineered receptor that change shape...concept, except the ligand is attached to a protein binding partner or nanobody that targets the native receptor... when exposed to light, the ligand engages the binding pocket and activates the receptor. Created with...absence of light the LOV domain blocks the enzyme binding domain preventing downstream signaling pathways...

  34. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    Published
    June 1, 2023, 1:15 p.m.
    ...experiment. Some inducible constructs can exhibit leaky expression in the absence of the inducing drug, so be...different ways to ultimately achieve success, we find it is usually faster and easier to do the validation...it can be ubiquitous, cell-type selective, or inducible.  Ubiquitous promoters  These often-strong promoters...over time, resulting in decreased expression (Damdindorj et al., 2014; Nieuwenhuis et al., 2021). It is... Attribution 4.0 International License.   Inducible promoters  This type of promoter allows you to... turn off, gene expression by administering an inducing agent like doxycycline (Favre et al., 2002). They...experimental output. You’ll also want to keep in mind the serotype you’ve picked, since that can affect...

  35. Human Germline Editing Using CRISPR

    Type
    Blog Post
    Published
    Aug. 10, 2017, 2:19 p.m.
    ...controls.) 27.6% of embryos carried indels at the mutation site, indicating that repair proceeded through NHEJ...move forward from this work. Experiments & major findings Let’s start with the paper itself: Ma et al. were...electroporated them with an ssODN repair template in induced pluripotent stem cells made from the patient. With...allowing repair to take place before DNA replication. Indeed, they observed that 41/42 embryos were non-mosaic...using the provided ssODN template. In fact, this finding led the authors to title the paper “Correction ...pre-implantation genetic diagnosis (PGD) screens embryos to find those that do not carry a given dominant disease-causing...pregnancy, the researchers were able to examine each individual blastomere to determine its genotype. However...

  36. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    Published
    Dec. 16, 2015, 3:30 p.m.
    ...on the Addgene Website Find CRISPR Plasmids Browse our CRISPR Guide Pages Find Genome Engineering Resources...combined with the weak separation afforded by gRNA binding at non-target sites would theoretically decrease... Figure 1: Strand separation and target DNA binding to the Cas9 nuclease. Stable strand separation ...similarly thought that if they could weaken sequence independent interactions between Cas9 and DNA, then they ...SpCas9-HF1 each stall in an inactive form after binding to a target site and propose that this stalling...Science (2015): aad5227. PubMed PMID: 26628643. Find plasmids from this publication at Addgene. 2. Nishimasu...effects." Nature (2016).  PubMed PMID: 26735016. Find plasmids from this publication at Addgene. 5. Chen...

  37. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...Screening Kit Find Out How to Best Utilize Our Cancer Resource Pages  Resources at Addgene Find the ClonTracer...exciting new tool that allows researchers to individually label millions of mammalian cells through lentiviral...Novartis because she wanted first-hand experience in industry research. When she came to Novartis, she was happily...these groups with the same drug regimen, they would find one of two outcomes (see schematic depiction below...ClonTracer library allows a researcher to label individual members of a starting population of cells with...multiplicity of infection (MOI) such that each individual cell receives only one barcode. The abundance...is a “really great way of broadly sharing these kinds of reagents.” Here at Addgene, we’re excited to ...

  38. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ...: PMC49604. 4. Independent control of immunoglobulin switch recombination at individual switch regions...PMID: 8513499. Resources on Addgene.org Find Cre plasmids Find loxP plasmids  ...-lox system is a technology that can be used to induce site-specific recombination events. The system ...key advantage of the system is that Cre acts independently of any other accessory proteins or co-factors...recognition sequences consisting of two 13-bp long palindromic repeats separated by an 8-bp long asymmetric ...certain stages of development, or by making the Cre inducible (such as with tamoxifen or doxycycline), the Cre...unique asymmetric spacer "NNNTANNN", where "N" indicates which bases may vary from the canonical sequence...

  39. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ...Learn about Addgene's viral service Find viral protocols and videos Find AAVs for your research ...historical ties to psychology and philosophy (the mind-body problem). Because of the availability of tools...between animals as olfactory stimuli that were left behind from a preceding animal (ex: urine) can be distracting...Animal Strain, age, sex, housing conditions (individually versus in groups), health status, diet, cage...differences in behavior observation); experimenter blinding to avoid bias Genetic manipulation in behavioral... neuronal targeting after stereotaxic injection. Find more details about choosing AAV or lentiviral vectors...about two weeks before the behavior test. This time window between injection and testing will vary depending...

  40. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    Published
    July 18, 2019, 12:55 p.m.
    ... International C. elegans meeting, I was again reminded of the extent that C. elegans researchers embrace...worm community where one can publish brief novel findings. These results are normally one figure and can...This a great resource for scientists to publish findings that don't fit into larger publications and would...embryo when all essential embryonic genes are individually depleted via RNA interference (RNAi). Scientists...tools for the community. Plasmids for the Auxin Inducible Degradation System (AID) The Dernberg lab in 2015...controls the expression of a GAL-4 driver that can bind to activation sequences upstream of genes to stimulate...expanded on this tool by splitting cGAL4 in two and binding each half to a gp41-1-N-intein. Split inteins are...
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