...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...

...helix. The order of these 4 nucleotides makes up the genetic code and provides the instructions to make ...Scientists often “sequence DNA” to identify the order of these four nucleotide bases in a particular DNA strand... protocols and videos for specific techniques and tips! Addgene Protocols Addgene Video Library...Addgene's molecular biology reference guide, including information on molecular genetics, plasmids, sequencing... the conclusions that genes encode proteins and reside on chromosomes, which are composed of DNA. These... transcribed from DNA. DNA is comprised of 4 nucleotides or bases, adenine, thymine, cytosine, and guanine...amino acids. Each amino acid is encoded for by 3 nucleotides termed a codon. As there are only 20 natural ...

... Science Guides CRISPR Guide Plan Your Experiment Plan Your CRISPR Experiment Jump To: Plan Your Experiment...selection marker to identify and enrich positive cells Packaging and envelope plasmids provide the necessary...see our Lentivirus Guide ) Stable, tunable expression of Cas9 and/or gRNA in a wide variety of mammalian... desired edit, ideally less than 10 bp away. In this case, it is necessary to identify the exact location... increase specificity, you can also consider using a high-fidelity Cas enzyme. In addition to off-target...the genome like never before. This section will provide a general framework to get you started using CRISPR...apply to using CRISPR in other organisms. First, consider the genetic manipulation that is necessary to ...

..... Cloning Resources Empty Backbone Guide Genome Engineering Guide Restriction Enzyme Gateway TOPO Gibson...Addgene's reference guide on molecular cloning techniques, including information on seven commonly used...is likely that for any given project there is an ideal approach. This may be due to speed, cost, availability...method for your research project . The following guide will highlight several of the most popular cloning...DNA that already has restriction sites on either side of it into any plasmid that has the same sites in...sequence, which can make it difficult at times to identify compatible restriction enzymes, that cut your ...polymerases synthesize DNA molecules using the 4 nucleotides and lastly DNA ligase fuses DNA strands together...

...Fluorescent Proteins 101 Science Career Guide Videos Watch our collection of video content , including protocols...eBooks, science guides, videos, and protocols Educational...Techniques Protocols Find written and video protocols for a wide range of applications in molecular biology... Educational Resources From eBooks to videos and blog posts... to protocols, our educational resources cover a wide range of topics in molecular biology, laboratory...Creating Bacterial Glycerol Stocks Restriction Digests Guides Browse our in-depth instructional content on subjects...

...Select Science Career Guide November 2018 | First Edition About Science Career Guide November 2018 |...your gRNA and how to use pooled libraries in genome-wide screens. Close About Overlay Select Fluorescent ...Vectors 101 July 2024 | Second Edition About This guide to viral vectors contains an introduction to each... | First Edition About The guide includes information on the many career options in science, advice ...advice on mentoring and management, resources for identifying your transferable skills, and tips on the job...

... are located within promoter regions, and they provide a stable binding site for RNA polymerase and transcription...Pribnow box which usually consists of the six nucleotides, TATAAT. Proximal Promoter Further upstream from...prokaryotic promoters. Eukaryotic promoters span a wide range of DNA sequences. It is not unusual to have...

...Primers? The primer sequences listed on the left are provided for your reference. Addgene does not distribute...primers based on the plasmid backbone and insert. To identify primers that may be useful in your sequencing ...