... 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...

...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...

... Science Guides Adenoviral Guide Adenoviral Guide...Guide to adenoviral components and vector construction Science...i nverted t erminal r epeats (ITRs). Genes are divided into early (E1-4) and late (L1-5) transcripts. ...the adenoviral plasmid. Please see our Biosafety guide for more information on working with adenovirus....replication. For more information about AAV, read our AAV guide . Can I make a stable cell line with adenovirus?...deleted from the shuttle vector. Early gene E1 is provided by the transfected cell line (either 293 or 911...generation of recombinant adenoviruses: A Practical Guide for using the AdEasy™ System (127 KB) . Glossary...

... 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...

...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) ...

... Guides Lentiviral Guide Lentiviral...systems are considered safer than 2nd generation systems because the packaging vector has been divided into ...and should be considered on a case by case basis. Biosafety should always be considered with respect to...Guide to lentiviral components, generations, and safe use in the lab Guides...Lentiviral Guide Components 2nd Generation 3rd Generation Viral Production FAQ References Glossary Lentiviral...planning your experiment, the important component to consider and optimize is the transfer plasmid. 2nd generation...(not pictured). Unless an internal promoter is provided, gene expression is driven by the 5'LTR, which...

... 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 ...

... Guides γ-Retrovirus Guide γ-Retrovirus...infectivity): The use of the VSV-G envelope provides the widest tropism or range of cells a virus can infect...Guide to retroviral packaging systems and commonly asked questions Guides...γ-Retrovirus Guide Packaging Systems Frequently Asked Questions (FAQ) Glossary γ-Retroviral Packaging Systems...Systems The widespread use of γ-retroviruses (gamma-retroviruses) in cancer and stem cell research has prompted...into the 293T packaging cell line. This system provides the greatest flexibility to pseudotype γ-retrovirus...information about lentivirus, please read our lentivirus guide. Are γ-retroviruses replication incompetent? Yes...

... 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...

... 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...

...AAV Guide Adeno-associated Virus (AAV) Guide Components Common Uses Viral Integration Serotypes Pseudotyping...Available at Addgene Biosafety Guide Parts of an AAV Transfer Plasmid Video Web References Vector Biolabs... AAV Guide will teach you about AAV components, serotypes, pseudotyping, and more. Science... Science Guides AAV ...Serotypes Eleven serotypes of AAV have thus far been identified, with the best characterized and most commonly...efficiently transduced by AAV2/2, and is distributed more widely in the brain, indicating improved transduction ...variants include: Self-complementary AAV (scAAV): One downside of AAV is its single-stranded DNA genome. Because...

...“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...

...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 ...

...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...

...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...

...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.   Fig. 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...

... 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...

...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 ...

...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...

...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...