Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 1,100 results for: Ide

Showing: 201 - 240 of 1100 results

  1. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...This is the next step in high-throughput, genome-wide screening,” Konermann said. “If you want to activate...only do that screen with a system you are really confident about working for most genes and individual targeting...protein. SAM can be combined with a human genome-wide library to activate all 23,430 known coding isoforms... we could use information from the structure to guide and improve activator design,” she said. “Before...exposed RNA loops, which Konermann recognized as an ideal spot to fuse the activators instead of at the protein...consistency and validation with >80% effectively enriched guides per gene hit, and 100% validation of the top 10...genes of interest. They also “showed that different guide RNAs have different abilities to turn genes on and...

  2. CRISPR-Cas14: a family of small DNA-targeting enzymes enabling high-fidelity SNP genotyping

    Type
    Blog Post
    ...enzymes." Science 362.6416 (2018): 839-842. PubMed ID PMID: 30337455. Additional resources on the Addgene...systems were merely adaptive immune systems that provide bacteria protection against infectious agents. ...predicted that many others are still unknown. Identifying Cas14 variants In this quest to discover new ...that improved specificity of Cas14 enables high-fidelity SNP genotyping. The development of  Cas14-DETECTR...Find the Cas14 plasmids here Checkout our CRISPR guide page Browse the CRISPR collection ...

  4. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ...in the apo state (PDB ID 4CMP, top) compared to the guide RNA-bound state (PDB ID 4ZT0, bottom), seen from...structure (left, PDB ID 4ZT0) with Cas9:RNA:DNA electron microscopy structure (right, PDB ID 7S4X), viewed from...bound to guide RNA (PDB ID 4ZT0), shown opaque (left) and transparent (right). The RNA guide sequence ...Figure 5: S. pyogenes Cas9 bound to guide RNA and target DNA (PDB ID 7S4X). Residues R1333 and R1335 from...structure of S. pyogenes Cas9 bound to guide RNA and target DNA (PDB ID 7S4X), shown opaque (left) and transparent...Figure 7: S. pyogenes Cas9 bound to guide RNA and target DNA (PDB ID 7S4X). The end of the DNA:RNA heteroduplex...structure of S. pyogenes Cas9 in the apo state (PDB ID 4CMP) with domains colored as in Figure 1. HNH, red...

  5. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    Blog Post
    ...with the Plasmid Name, Addgene Plasmid ID #, and the PubMed ID # (PMID) for the article that has used ...it has been validated in.  If a plasmid's Addgene ID # has been referenced in other publications, you'...used your plasmid of interest, which may help you identify relevant protocols and experimental data Find...

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

    Type
    Blog Post
    ...we request that you cite it using both its Addgene ID number as well as the reference for its original ...cite a plasmid from Addgene using its unique Addgene ID number, your publication will be listed on our website... You can search for plasmids by name, PI, Addgene ID number, or keyword. Although an initial search might...plasmids from Addgene right on our plasmid pages and provide a citation that you can put straight into your ...search results using the filters found on the right side of the search page. A great breakdown of our search...search functionality can be found in the following video: Please remember that the accuracy and reliability... Blog Post on Our Citations Feature Browse More Inside Addgene Posts Additional Resources on Addgene....

  7. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    ...were able to include specific plasmid ID numbers. If you know the ID number for a plasmid, your straightforward...by our own CSO. But what if you don’t have plasmid ID numbers, or you aren’t sure if any plasmids from ... associated plasmids through the article’s PubMed ID (PMID) ...or DOI Or get some keywords close enough...more. Selecting crispr in Guides will return links including our CRISPR guide.  For a closer look at this...search. These pages provide helpful information about the subject, link to related guides and protocols, and...delivering a calcium biosensor using AAV, you might decide you’d really prefer to order ready-to-use virus...search will start from a publication. You may have an idea for a plasmid that would be useful in an experiment...

  8. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...entering the data. This step associates an Addgene ID with your plasmids, and adding all the plasmids at...likelihood that the plasmids are given sequential IDs.   Name – we encourage the use of descriptive plasmid...include the total size of the insert and the GenBank ID associated with your insert. Pro Tip! If the correct...and the vector type. Pro Tip! Reference the Addgene ID in the "Backbone Manufacturer" box for backbones ... Confirmation email from us with a unique Deposit ID. Addgene will send you a package containing instructions...scientists worldwide. What's more, depositing is free! We've written this post as a step-by-step guide to the...will not make the plasmids available until you provide us with updated publication information or permission...

  9. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    ...Addgene plasmid ID (if we have one associated with the gRNA), application, cas9 species, PubMed ID, and depositor...published article. This blog post will serve as a guide to use our validated gRNA Target Sequence Table....sequence variations (e.g. SNPs) exist. Additionally, consider testing multiple gRNAs targeted to the same locus.... As you develop and confirm new gRNAs, please consider submitting their sequences (and plasmids!) so ...Addgene.org Browse gRNA Design Tools Read Our CRISPR Guide Pages Find CRISPR Plasmids  ...

  10. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...and make sure they have been barcoded with a unique ID number (1D barcode as shown below). This number ensures...samples. They will help your lab ensure that sample ID’s are unique, consistently labeled, and that numbers...barcode to the container holding the tubes and use this ID to name the corresponding Excel sheet.3. Be specific...the 2D barcodes mentioned above. These barcodes identify what type of sample is contained within the tube...tube’s appropriate location. The barcode also identifies the contents of each individual tube.  If a plate...bacteria with the plasmid can grow. As the bacteria divide, they replicate and retain the plasmid. This allows...store a lot of information in a small space and is ideal for tracking large collections. Many companies, ...

  11. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    ...search scoped to that category. Catalog ID suggestions provide shortcuts to specific catalog items as ...contains a plasmid with the catalog ID). Depositor suggestions provide shortcuts to matching PI labs and...educational resources — including blog posts, protocols, guides, and collections — is a lot of material to sift... the different usage patterns the larger team identified. The basic function of a search engine is to ...through our educational resources. Filters We identified the most useful kinds of information per resource...words, phrases, partial phrases, or numerical identifiers. We also combine the different terms in a query... name in the publication field. Suggestions We provide as-you-type suggestions in different ways. Keyword...

  12. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ...the former, you simply type in identifier keywords (Cas name, RefSeq ID, etc.) and you will see a table...got you covered!     Fig. 1 – CasPEDIA Cas ID display information for a given Cas (SpyCas9a) from...not so long ago, spCas9 was the only Cas enzyme widely available and applied by researchers for gene targeting...experimental fit due to this. But now, it’s time to cast aside those concerns and embrace a new resource to solve...CasPEDIA is an online, searchable resource that provides summary information on Class 2 Cas enzymes. The...requirement, gRNA, and multiplex ability Experimental guidelines and applications Protein structure and properties...experiment. Target requirements As we all know, a Cas guided by a gRNA is useless without an adjacent PAM or...

  13. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...with the ID of the component plasmid, component name, size, and any comments. You can use the ID to look...it’s easy to switch out reporters, Cas proteins, guide cassettes, promoters, N’ and/or C’ terminus, and...Bbsl, a Type II restriction enzyme that cleaves outside of its recognition sequence. This means that each...Mammalian Lentivirus pRDA_512 (RSV-driven; lentiGuide-based) Mammalian Lentivirus pRDA_722... the design process down into four easy steps, provides the real value for the user. Let's walk through... Plasmids 101: Modular Cloning Addgene's CRISPR Guide Plasmids 101: Golden Gate Cloning...

  14. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ... Addgene ID numbers and the reprogramming factors they encode Episomal Plasmid Addgene ID# Encodes ...iPSCs. The generation of human iPSCs (hiPSCs) now provides an opportunity to develop and use patient-specific...fibroblasts [3]. These advantages make PBMCs the ideal source of somatic cells for reprogramming purposes...field [4]. Three major reprogramming methods are considered when generating hiPSCs for clinical purposes:... of the procedure. Episomal reprogramming is an ideal and cost-effective method for creating clinical-...reported efforts. The pCXLE episomal system thus provides a practical way to generate transgene-free, virus-free...

  15. Science Guides

    Type
    Guide
    ...Collection of guides that describe how popular scientific techniques work... Science Guides Addgene has created... created the following guides to help you start learning about various scientific concepts related to ...to our plasmid collection. Find links to the guides below. Antibodies Chemogenetics CRISPR Optogenetics...molecule, can be used to capture, quantify, and identify proteins of interest. Read More Chemogenetics ... sets. Read More Viral Vectors Find educational guides to learn more about the different types of viruses...

  16. Adenovirus Guide

    Type
    Guide
    ...Adenoviral Vector Guide Adenoviral Vector Guide Jump To: Adenoviral Vectors Vector Production Integration Adenoviral...homologs, especially when having to express them alongside a guide RNA and other regulatory elements. The packaging...production, in trans refers to genetic elements provided outside of the plasmid containing the gene of interest... vector guide to learn about adenoviral vector components, production and common uses. Guides... Guides Adenoviral...a protein capsid and the viral genetic material inside. Adenoviral genomes range between 26–45 kb in length... terminal repeats (ITRs). Adenoviral genes are divided into early (E1A, E1B, E2, E3, E4) and late (L1,...

  17. Chemogenetics Guide

    Type
    Guide
    ... Science Guides Chemogenetics Guide Chemogenetics Guide Jump To: Chemogenetics Guide Early Chemogenetic...) is a widely-used tool for achieving in vivo expression of chemogenetic receptors, and a wide variety...and this must be considered. For tissue specificity of AAV serotypes, see our AAV guide . FLEx Vectors ...Addgene's guide to using Chemogenetics plasmids in your lab for interrogation of neuronal activity. Science...the application of a small molecule ligand. The ideal chemogenetic tools are unresponsive to native ligands...signaling. GPCRs have been shown to be involved in a wide variety of biological processes, including initiating...of constitutive activity, making them less than ideal. DREADDs Designer Receptors Exclusively Activated...

  18. CRISPR Guide

    Type
    Guide
    ...Science Guides CRISPR Guide CRISPR Guide Jump To: CRISPR Guide Overview CRISPR Basics Small Edits Large...making CRISPR perfect for genome-wide screens. This guide will provide a basic understanding of CRISPR ...genomic target of the gRNA can be any ~20 nucleotide sequence, provided it meets two conditions: The sequence...discuss more Cas9 fusion tools later in the guide. Increasing fidelity of Cas9 Researchers have also enhanced...Cas9 into high fidelity Cas9s (hfCas9) . There is currently no defined measure of fidelity, so sometimes...with higher fidelity than SpCas9. FrCas9 (from Faecalibaculum rodentium ) has increased fidelity, and recognizes...some base editing systems create a wide spectrum of single-nucleotide variants (somatic hypermutation) ...

  19. Optogenetics Guide

    Type
    Guide
    ... Science Guides Optogenetics Guide Optogenetics Guide Jump To: Optogenetics Guide Introduction Microbial...have been created and identified in other species - by acting as light-gated chloride channels, these variants... mutations 540 Channelrhodopsins: chloride channels iChloC Chloride-conducting channel, CrChR2 with mutations...Addgene's guide to using Optogenetics plasmids in your lab, including explanations of microbial opsin...ions into or out of the cell. Scientists have identified a variety of naturally occurring microbial opsins...stimulated. Channelrhodopsin-2 (ChR2), the first widely adopted optogenetic tool, is also a blue light ...been expanded by scientists through both the identification of novel ChRs from other algal species and ...

  20. Antibody Guide

    Type
    Guide
    ... Science Guides Antibody Guide Antibody Guide View Addgene's Antibody Collection...This method uses peptide identification by proteomics (Capture MS) to validate the identity of proteins recognized...first identify your assay type and what species your protein of interest comes from. Then consider the ...In our Antibody Guide, we define key terms, summarize antibody production and storage techniques, and...Collection Jump To: Antibody Guide Production Structure Isotypes Antigen Specificity Other Affinity Reagents...Antibodies can be used in the lab to quantify, identify, separate, and locate proteins of interest. The... between antibodies, allows for the antibody’s identity to be easily confirmed via sequencing, and enables...

  21. Lentiviral Vector Guide

    Type
    Guide
    ... Guides Lentiviral Vector Guide Lentiviral Vector Guide Jump To: Lentiviral Vectors Lentiviral...production, in trans refers to genetic elements provided outside of the plasmid containing the gene of interest...Read our lentiviral guide to learn about lentiviral components, generations, lentiviral production, and...mutations, or treatments into cells, and has been widely used in the research community for decades. Wild-type...in a host, and exhibit low immunogenicity. This guide contains many viral vector-specific terms and acronyms...gamma-retroviruses, see Addgene's gamma-retroviral vector guide . Lentiviral Vectors The genome of lentiviruses ...contains packaging gene env (usually VSV-G due to wide infectivity) VSV-G (vesicular stomatitis virus G...

  22. Guide to Using Pooled Libraries

    Type
    Guide
    ... Science Guides Guide to Plasmid Pooled Libraries Plasmid Pooled Libraries...amplification protocol. Read our CRISPR guide for more information on genome-wide screening with CRISPR/Cas9 . Browse... Libraries Jump To: Pooled Library Guide Introduction Library Amplification Types of Screens Resources...powerful tool for forward genetic screening, or identifying previously unknown genes that contribute to a...replicate for that plasmid. Library screens can be divided into two types: positive screens and negative screens...septicum alpha-toxin , and resistance to to the nucleotide analog 6-thioguanine . Next-generation sequencing...Positive Screen In a positive screen, the goal is to identify those cells that survive post-selection. The selective...

  23. Gamma-Retroviral Vector Guide

    Type
    Guide
    ... Guides Gamma-Retroviral Vector Guide Gamma-Retroviral Vector Guide Jump To: Gamma-Retroviral...production, in trans refers to genetic elements provided outside of the plasmid containing the gene of interest...Read our gamma-retroviral guide to learn about gamma-retroviral components, retroviral production, and...mutations, or treatments into cells, and has been widely used in the research community for decades. Wildtype...expression in a host and low immunogenicity. This guide contains many viral vector-specific terms and acronyms...about lentiviruses, see Addgene's lentiviral vector guide . Gamma-Retroviral Vectors The genome of gamma-retroviruses...contains packaging gene env ; usually VSV-G due to wide infectivity VSV-G (vesicular stomatitis virus G ...

  24. We're Updating Our Hold for Publication Policy

    Type
    Blog Post
    ...addgene.org with the citation and your deposit or plasmid ID numbers and we’ll be happy to update the plasmids...submit plasmids at any point in that process and we provide an option to keep your deposit on hold from public... their materials as soon as possible, whether alongside a journal article, a preprint, or without any ...openly sharing materials are both excellent ways to provide transparency, support reproducibility, and accelerate...with journal articles. But we do encourage you to consider sharing them openly as soon as that preprint is...

  25. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...Vector Guide Adeno-Associated Viral (AAV) Vector Guide Jump To: AAV Vectors Vector Production Integration AAV...watching our Optogenetics 101 video or by visiting Addgene's Optogenetics Guide . Browse optogenetics plasmids...production, in trans refers to genetic elements provided outside of the plasmid containing the gene of interest...viral vector guide to learn about AAV vector components, production and common uses. Guides... Guides AAV Vector ...a protein capsid and the viral genetic material inside. The AAV genome is ~4.7 kb long and consists of...viral vectors for gene therapy applications. This guide contains many viral vector-specific terms and acronyms...

  26. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

    Type
    Blog Post
    ...“plasmid” derived as a hybrid of "cytoplasm" and "id" (Latin for 'it'), as “a generic term for any extrachromsomal...noted that, unlike F-factors, the evidence did not support the idea that R-factors could integrate into... Élie Jacob and François Wollman and became the widely adopted name for these elements. At the time, the...Herbert Boyer, and Charles Brinton concocted a wild idea of using the newly discovered EcoRI enzyme (and ...such as pBR322, pACYC, and pUC were developed to provide higher copy number vectors that could be used in... with our goal of creating a one-stop reference guide for plasmids we've updated and added new content...Datta, Naomi. “Transmissible Drug Resistance in an Epidemic Strain of Salmonella Typhimurium.” The Journal...

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

    Type
    Blog Post
    ...ssODN: Using Ensembl Transcript ID(s) as an input query, pick crRNA (guide RNA sequence targeting the gene...tagging experiments: Retrieve Ensembl Transcript ID(s) of your favourite gene(s) from Ensembl genome ...coupled to high-throughput sequencing to identify genome-wide binding patterns of the target protein. ... not work in pull-down assays: Epitope tagging provides a solution Proteins - the workhorses of the cell...complexes are then analysed by mass spectrometry to identify the interacting proteins. Similarly, in ChIP-seq.... Both IP/MS and ChIP-seq methods provide a wealth of information about how protein networks work in ...to mention frustrating. One elegant solution to sidestep the antibody problem is to tag the protein of ...

  28. Dos & Don'ts When Publishing a Scientific Manuscript

    Type
    Blog Post
    ...bit like a GenBank ID) to all antibodies, animal models, and software tools in the Materials and methods...scope for the journal’s coverage. These guidelines also provide information on a journal’s house style ...Resource Identification Initiative. During this project, authors are asked to add an identifier (a bit ...vivo research and the CARE guidelines for case reports are two sets of guidelines frequently used by editors...information is provided. The clearer you write your paper and the more information you provide on data, materials...publication in a scientific journal requires several considerations. First, it is important to establish the type...audience you want to reach with your paper. Is it the wider scientific community, or are you writing for a specific...

  29. Summer SciComm: Preprints

    Type
    Blog Post
    ...significantly sooner. Plus, authors can include the Addgene ID for their deposited plasmids in the final journal...used in a preprint provides all the same benefits as sharing those plasmids alongside a journal article...different goals, formats, and processes, and they provide different amounts of access or feedback. One of...in the past few years.   Figure 1: Stand aside, CERN — it's a new age of accelerating science! ...contrast, journal-based peer review is usually a confidential process among a few individuals.  Early-career...especially enjoy the ability to post a preprint as evidence of their productivity that can be shared with ...funding bodies and review committees encourage and consider preprints as demonstration of productivity. Part...

  30. Reproducibility for Everyone: Lessons from an Open Science Collaboration

    Type
    Blog Post
    ...Reproducibility Read a blog post on Unique and persistent IDs for improved reproducibility Other resources: Reproducibility...practices could be an actionable way to tackle widespread irreproducibility. Luckily, there were other ...academics and like-minded organizations with similar ideas who were also thinking about reproducibility and...Thanks to Teytelman’s vast network and ability to identify the right opportunities, he was able to bring ...and motivated groups and individuals with similar ideas to actually do something about it. Building a diverse...received, and then when the team started brainstorming ideas for the next workshop, they realized that holding...enable reproducibility practices being adopted by a wider group of researchers. Generating material to empower...

  31. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    ... a post-doc in the Target ID and Validation Group and says that she decided to do a post-doc at Novartis...library, propagated it, separated it into roughly identical replicate groups, and treated these groups with...cell line containing an activating mutation in epidermal growth factor receptor (EGFR) with the ClonTracer...models. She hopes that the library can be used to identify appropriate, non-overlapping targets for drug ...distinct evolutionary paths to become resistant to epidermal growth factor receptor inhibition." Nature medicine...

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

    Type
    Blog Post
    ...Looking at the structure of the Cas9 nuclease (PDB ID: 4OO8 and 4UN3), Slaymaker et al. hypothesized that...Benjamin P., et al. "High-fidelity CRISPR–Cas9 nucleases with no detectable genome-wide off-target effects."...As evidenced by all the CRISPR publications, press, and plasmids out there, it’s obvious that CRISPR ...1.1), followed by a technique for detecting genome-wide double strand breaks (BLESS), revealed that these...as predicted, decrease off-target effects genome-wide. Further work by Slaymaker et al. showed that these... al. "Crystal structure of Cas9 in complex with guide RNA and target DNA." Cell 156.5 (2014): 935-949....Website Find CRISPR Plasmids Browse our CRISPR Guide Pages Find Genome Engineering Resources ...

  33. Lab Automation at Addgene

    Type
    Blog Post
    ...removable parts to fit on the deck of the Biomek. The FXP id also able to do high throughput DNA preps and other...purification, and filling vials with agar. I’ll also provide you with some tips on how to automate some simple...protocol and, after looking at a few options, we decided on Beckman Coulter’s Biomek FXP. We had a previous...scale. Using the BIOROBOT Universal Platform is ideal because it eliminates the potential of human error...standard molecular biology techniques Browse our video library Check out our molecular biology reference...

  34. Common Injection Routes in Mice

    Type
    Blog Post
    ...Skill required: beginner Intradermal Intradermal (ID) injections occur between the layers of the skin.... post has provided a good introduction to common injection routes in mice! When considering what method...administering vaccines, pain meds, or other drugs. Identifying the proper injection site is straightforward,...knowledge. Intrathecal injections are used for a wide array of purposes, but mainly for the delivery of...sounds very disruptive, but surprisingly, it is considered minimally invasive. Restraint: anesthesia Location...common. Due to the intubation, this procedure is considered quite invasive. Restraint: anesthesia Location...feeding tube with a bulb on the end, which prevents accidental tears or punctures to the throat. The feeding...

  35. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ... It’s possible to associate the depositor’s ORCID ID with the deposit, and a DOI is added to the deposit...integration is described in this video and the Figshare integration in this video.  The ability to link from...making it searchable and shareable, but some ELNs provide better support for reproducibility than others....reproducibility is limited in two ways – only data inside the ELN is reproducible, and only the highly restricted...requirements that emerged from intensive engagement with a wide range of people at the University of Wisconsin in...principles in mind, and in response to further input provided by the research communities at the University ...online file storage tools, as illustrated in this video, and to data stored on institutional and lab servers...

  36. Tips for arabidopsis transformation

    Type
    Blog Post
    ...https://books.google.at/books/about/Arabidopsis.html?id=IfZAMNPWVk4C&redir_esc=y (Accessed: 21 September ...culture into a container that is wide enough to dunk your plant’s flowers inside. 50 mL falcon tubes or plastic...process right the first time. I’ll lay out some guidelines here. A detailed transformation protocol, as ... et al. (2006) ‘A rapid and robust method of identifying transformed Arabidopsis thaliana seedlings following...

  37. The Challenges of Cell Culture

    Type
    Blog Post
    ...Short Tandem Repeat (STR) profiling can be used to ID the cell line. The same test is so good that it is...many journals strictly require evidence of cell line provenance and identity. The Nature Publishing Group...Resources ATCC Animal Cell Culture Guide Thermofisher Guidelines for Maintaining Cultured Cells Invitrogen...Invitrogen Cell Culture Basics Guide Abcam Cell Culture Guidelines Resources on the Addgene Blog Learn ...culture in the present day Cell culture is a widespread tool used in the fields of oncology, virology...this reason. Unreproducible results are rife. Misidentified or cross-contaminated cells have been known ...Invasion in Renal Cancer Cells, Marker-independent identification of glioma-initiating cells. The International...

  38. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...desired insert (e.g., mScarlet).   Plasmid ID Plasmid Name Tags 69090 pMJ915 MBP (N...differences in guide RNA efficiency. For example, in a two-guide floxed experiment, if one guide cleaves less...from the target exon on either side. They have successfully used guides targeting the same or complementary...on each side. For optimal repair, place the loxP sites precisely at the cut site for each guide. This design...RNP) complexes of Cas9 protein and the separated guide RNA components (crRNA and tracrRNA) for maximum ...given location, you’ll need to design only a single guide and your donor ssDNA. Here, we’ll use the example...homology-directed repair experiments, you’ll want the guide cut site to be as close as possible to your desired...

  39. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...Biology. New York: W. H. Freeman, 2000. Bookshelf ID: NBK21668. Matsuno-Yagi, Akemi, and Yasuo Mukohata...neuronal subpopulations, and this represents a major downside in optogenetics (Belzung et al., 2014). This could...genes. Strong cell-specific promoters, on the other side, are too long for rAAVs, which can only package ...Double-floxed Orientation)) is a powerful tool that provides precise temporal and spatial control of gene expression...produce behavioral and electroencephalography evidences of arousal in mice previously subjected to steady-state...., 2019). FLEx vectors represent, therefore, an ideal partner to optogenetics to understand the cellular...

  40. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    .... (2024). A guide to selecting high-performing antibodies for Synaptotagmin-1 (Uniprot ID P21579) for ...antibodies when possible. If cost is prohibitive, consider an approach that prioritizes more reproducible... rate. If you need a more permanent knockdown, consider using shRNA vectors instead. With both siRNA and...this approach should also allow for the clear identification of the cell types of interest. By correlating...correlating your data sets, you can increase confidence that the labeling observed represents the expression ... at similar levels in your cell lines, you can consider a cell treatment approach. If there is a cell ...protein. A challenge to this approach is that the identity of the epitope is often not available to confirm...
Showing: 201 - 240 of 1100 results