...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ... or want to swap out the constant domains, the plasmids for the heavy chain and light chain are also available...ubiquitination and degradation.   Find the CRBNmidi plasmid here!   Kroupova, A. et al. (2024). Design of ...minimal background.   Find these C. elegans mScarlet plasmids here!   Cao W.X., Merritt D.M., Pe K., Cesar ...disruption in Mycobacterium abscessus using a dual-plasmid CRISPR/Cas9 system By Aryana Khosravani Deborah...Deborah Hung’s lab has developed a novel dual-plasmid CRISPR/Cas9 system for gene disruption in Mycobacterium...methods). By using mCherry for selection of sgRNA plasmid transformants, they avoid M. abscessus’ high rate...

...of the new plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles. ...be used for applications beyond cryo-EM.   Find plasmids for MagIC-Cryo-EM here!   Arimura, Y., Konishi...disease phenotype in an animal model. Find PE-eVLP plasmids here! The lab’s other recent deposit builds on...over the wild-type enzyme. Find optimized PASSIGE plasmids here!     Figure 3: Schematic of the PASSIGE...recombinase integrates a sequence from a donor plasmid containing a complementary site into the genome...without compromising their efficacy.   Find CHARM plasmids here!   Neumann, E. N., Bertozzi, T. M., Wu, E...

...of the new plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles.  ...2 nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane protein (Plasmid #141386-LV [+M], negative... over the original CROPseq. Find CROPseq-multi plasmids here Walton, R.T., Qin, Y., Blainey, P.C. (2024...potential improvements to the editor. Find PE7 plasmids here!     Figure 2: PE7 improves prime...conjugated with its HaloTag domain.  Find Voltron2 plasmids here! Abdelfattah, A.S., et al. (2023) Neuron...therapeutic applications.  Find microglia-targeting AAV plasmids here! Serrano, C., et al. (2023). Simple and ...potential for therapeutic success. Find REVeRT plasmids here! Riedmayr, L.M., et al. (2023). mRNA trans-splicing...

...on Addgene.org CRISPR Plasmids – Protein Tagging CRISPaint Gene Tagging Plasmids 101 eBook Resources ...Resources on the Addgene blog Plasmids 101: Protein Tags Plasmids for Endogenous Gene Tagging in Human Cells ...cloned in-frame to the protein of interest for plasmid-based expression systems. Degron tags are one of...

...As a plasmid repository, we are no strangers to cloning. If you’re reading this, then chances are you...NEB’s DAD Interview with Nicola Patron on MoClo Plasmids 101: Golden Gate Cloning ...

...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...all applications. Addgene’s plasmid collection includes over 10,000 plasmids with a FLAG tag (!), but did... al. 2023 with permission.   Find Cas-PAGE plasmids here!   Zhang, Z., et al. (2023). Efficient engineering...inspired and informed this section.  Find mStayGold plasmids at Addgene here!   Ando, R., et al. (2023). StayGold...under CC-BY license.   Find the AAV.CAP-Mac plasmid here!   Chuapoco, M.R., Flytzanis, N.C., Goeden...(2023) under CC-BY license.   Find AAV1-X1 plasmid here! Chen, X., et al. (2023). Functional gene ...

... ship rabies plasmids and viral vectors containing genetic elements from rabies plasmids to international... materials in our repository, including rabies plasmids, are export-controlled materials in the USA, where...export license applications for VSV and SARS-COV-1 plasmids. Export control license process    An export ...license, once approved, allows Addgene to ship plasmids or viral vectors containing sequences from rabies...  Timelines  We have offered VSV and SARS-COV-1 plasmids under this application process since May 2020....at [email protected].  Curious about how rabies plasmids are used in the lab? Check out our “Rabies and...browse tools for AAV tracing, made from rabies plasmids deposited by Ian Wickersham’s lab.   ...

...of the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. ... missed something smokin'? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) ...Figure 3 Colognori et al 2022.   Find CRISPR Csm plasmids here! Colognori D, Trinidad M, Doudna JA. Precise...inducible expression.   Find heat-inducible lentiviral plasmids here!   Wu Y, et. al. Control of the activity ...2: Addgene’s western blot for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP... Campbell et al, Nature Methods   Find YFP plasmids here!   Campbell, B.C., et al. Chemically stable...

...Tet-Off system.  For more information on Addgene Tet plasmids, see our Tetracycline resource page. Cumate Switch...20697123. PubMed Central PMCID: PMC2942869. Find plasmids from this publication at Addgene. del Valle Rodriguez...20434990. PubMed Central PMCID: PMC2883883. Find plasmids from this publication at Addgene. Wei, Xing, ...22406855. PubMed Central PMCID: PMC3846601. Find plasmids from this publication at Addgene. Riabinina, ...25581800. PubMed Central PMCID: PMC4344399. Find plasmids from this publication at Addgene. Riabinina O...Hill, C. et al. A LexAop > UAS > QUAS trimeric plasmid to generate inducible and interconvertible Drosophila...

...of the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. ...Think we missed something hot? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) ... you can also find several other constructs in plasmid form. Fig. 1: Summary of the approach and...JEDI-2p viral preps here (and here!)Find JEDI-2p plasmids here!  Liu, Zhuohe et al. Sustained deep-tissue... improve the performance of HcKCRs. Find HcKCR plasmids here!   Govorunova EG, et al. Kalium channelrhodopsins...’ in the Addgene catalog pulled up over 13,000 plasmids! Our recombinant anti-c-Myc [9E10] (isotype IgG2a...myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the Applications section on the Anti-c-Myc...

...conjugative mobilization of broad host-range IncQ plasmids." Plasmid 62.2 (2009): 57-70. PubMed PMID: 19465049...page Deposit microbiology plasmids with Addgene Find synthetic biology plasmids ...family-RP4 conjugation system. This system can deliver plasmids to both Gram-positive and Gram-negative cells.... Wang lab further developed a modular suite of plasmids (pGT) with various replicative origins that can...genera across four phyla that have taken up the plasmid through conjugation in vivo studies. However, they... tools for microbiome studies.  Find the MAGIC plasmids here Paper-based synthetic biology approach to...conjugation. The labs screened for the host-range of the plasmids and suitability of various antibiotic resistance...

...to find Gateway plasmids for your organisms of interest: Find Gateway cloning plasmids here! References...antibiotic resistance markers (indicated here by plasmid color), allowing you to easily select for the expression...desired, recombined construct. Read our recent Plasmids 101 Post on CcdB for more information. How to ...Method A: recombination of an attB-PCR product or plasmid with an attP donor vector. In this case, we would... to use the Snapgene program to design Gateway plasmids.    Figure 3: Method A to create an entry...entry clones can be used with a large variety of plasmids. Speed The Gateway system enables the generation.... Take a look at some of the Gateway Multisite plasmids available at Addgene, including the Frew Lab Multiple...

...Deletions in a Plasmids Learn How to Verify Your Plasmid Resources on Addgene.org Find More Plasmid Cloning...

...the correct orientation or even if it’s in your plasmid backbone. Backbone-specific primers: A second...orientation of your insert or if the insert is in your plasmid Have to make a new primer pair for each insert ...about the size of the insert and if it's in the plasmid Can be used to screen clones that were created ...created with the same backbone because primers are plasmid dependent Doesn't provide information about the...insert Tells you if the insert is present in the plasmid Great for blunt end cloning You have to make...thermocycling program. One key difference is the plasmid DNA must be released from the bacteria in order... just need to lyse the bacteria to release the plasmid DNA by either briefly boiling the sample before...

...In a previous post from our Plasmids 101 series, we learned how the Cre-loxP recombination system can... even in the presence of Cre recombinase. This plasmid now specifically drives expression of mCherry instead...2286602. PubMed Central PMCID: PMC3406316. Find FLEx plasmids from these Addgene depositors: Ed Boyden Bernardo... plasmids... plasmid... plasmids 101... flex plasmid...

...previous posts for our Plasmids 101 series, we examined a number of important plasmid elements – promoters...Resources on Addgene.org Find Cre plasmids Find loxP plasmids  ... expression proceeds. One popular lox-stop-lox plasmid is from Tyler Jack's lab: Lox-Stop-Lox TOPO.  Cre-dependent...  plasmids 101...

...localization, or protease protection. Thus far Plasmids 101 has covered GFP and its related fluorescent...stability activity, can be readily purified using plasmids found in this paper (available at Addgene).   ...interest: Which Fluorescent Protein Should I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent...   plasmids 101...

...Browse our Plasmids 101 blog posts Read blog posts on plasmid cloning Read about different plasmid elements...to the backbone plasmid; a starting point could be a molar ratio of 1:2 (plasmid:insert), but this parameter...colonies screened might not contain the correct plasmid. There’s a lot that can go wrong! With all the ...cloning project, and hopefully obtain your coveted plasmid with no substantial delays. Design, design, design...spend some time outlining the construction of the plasmid and all the steps you will have to take. Decide...you chose, and verify that the sequence of the plasmid you are working with is correct. It's also best...ways you can assemble the different parts of a plasmid based on the cloning strategy you followed. While...

...collection of plasmids each carrying a single gene with a defined mutation or gene fusion. Plasmids carrying...cancer research, like PIK3CA (Plasmid 116771: pHAGE-PIK3CA) and EGFR (Plasmid 116731: pHAGE-EGFR), are represented...Resources on Addgene.org Find plasmids for your research Find cancer research plasmids and resources ...mutations, and the group has deposited several hundred plasmids from this study with Addgene. Read on to learn...learn more about their work, and how these plasmids can help cancer researchers studying an assortment of...well. They called this library the CTD2 FASMIC Plasmid Collection for “Functional Annotation of Somatic...cancer research community, but what else are these plasmids useful for? They could be used to more thoroughly...

...our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression...protein expression The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins at the beginning.... Now, we've turned some of the most requested plasmids into lentiviral preps to speed up COVID-19 research...Testing AAV inventory for the entire collection! Plasmid Serotype Name Depositor 27056 AAVrg pAAV-Ef1a-DIO...constructs that allow for targeted expression. Plasmid 75470 is now available in AAV1 for Flp-dependent... mCherry reporter. We now also offer AAVrg for plasmid 83898, which targets expression of ChR2 to forebrain...interneurons through the mDlx enhancer element. Plasmid Serotype Name Depositor 75470 AAV1 pAAV-CAG-FLEXFRT-ChR2...

...recombined plasmid to overtake full-size plasmids (or for smaller plasmids to overtake larger plasmids, if the...How many unique plasmids does your library contain?), the variance of size of plasmids in your library...lead to issues such as recombination and loss of plasmid diversity, so Addgene strives to distribute aliquots...library (How important is it that every single plasmid in your library is present, and how will you ensure...30 to 200 times as many colonies as individual plasmids in your library. As an example, if you want 100x... your library, and your library contains 5,000 plasmids, your goal for your transformation would be to...libraries often contain thousands, if not millions, of plasmids, they are generally electroporated into electrocompetent...

...features was our collection of popular plasmid sequences. These plasmids contain features such as antibiotic... our SnapGene powered plasmid maps Learn how we use SnapGene during the plasmid quality control process...out full plasmid sequencing at Addgene Additional Resources on Addgene.org Example plasmid analysis ...design was done painstakingly by hand. Records of plasmid construction were often incomplete or nonexistent...popular aspects - its ability to annotate a raw plasmid sequence and display frequently used genes and ...when the DNA sequence has changed. Identifying plasmid control elements Coding sequence features are relatively... boundaries for control elements in commercial plasmids were inconsistent and sometimes clearly wrong....

...is a global nonprofit plasmid repository. Over 2,000 labs have deposited plasmids to Addgene and we distribute...opportunity for Addgene to present our own data on plasmid deposits and distirbution for these fast moving...distribute over 130,000 plasmids in 2014. Thus, we are in a unique position to observe and quantify how...scientific community. Data on CRISPR and synBio plasmid sharing In a recent review on CRISPR technology...trends for Zinc Finger, TALE, CRISPR, and SynBio plasmids over time. You can see these data in the slide...molecular biology technology? Have you deposited your plasmids yet?      ...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... our hot plasmid articles from April 2018. If you'd ever like to write about a recent plasmid deposit ...sign up here. No time to read? Listen to our Hot Plasmids Podcast! TRIM-Away: Targeted endogenous protein...your protein knockdown experiments! The TRIM-Away plasmids can be found here. Clift et al. Cell. 2017. PubMed...Endosomal pH Sensors podcast segment The first Addgene plasmid deposit from Professor Stefan Hell, who won the...epigenetic memory at nearly any locus. The FIRE-Cas9 plasmids can be found here. Braun et al. Nat Communications...

...constructs from Christopher Harvey’s lab. These plasmids localize opsins to the soma to avoid off target...neighboring neurons. We have also packaged hChR2(H134R) plasmids from Karl Deisseroth’s lab, which includes pAAV-hSyn...H134R)-EYFP that is both Cre and Flp dependent. Plasmid Serotype Name 124603 AAV9 pAAV-EF1a-DIO-ChrimsonR-mRuby2... has now expanded to offer many of our control plasmids in new serotypes. See our Serotype Testing AAV...AAV inventory for the entire collection! Plasmid Serotype Name 51502 AAV5 pCAG-FLEX-EGFP-WPRE 114471... jGCaMP7 from Janelia, including three jGCaMP7 plasmids now packaged in AAVrg. jGCaMP7s, the variant with...post Overview of AAV Encoded Calcium Sensors). Plasmid Serotype Name 104489 AAVrg pGP-AAV-syn-jGCaMP7b-WPRE...

...CRISPR mediated genome engineering. 10. Plasmids 101: Control Plasmids Need help thinking about the types ...Scientists eBook Download the Plasmids 101 3rd Ed. eBook Read the Most Recent Plasmids 101, CRISPR 101, and Career... two eBooks (Management for Scientists and the Plasmids 101 2nd ed, the 3rd edition has now been released...top of the many developments in science and the plasmids, technologies, and researchers behind them. As...most popular posts continue to be those from our Plasmids 101 and CRISPR 101 series. These posts aim to ... the background they need to get started using plasmids and the revolutionary CRISPR technology in their...and how it’s used in CRISPR genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate...

...curated collections of plasmids as kits with greatly reduced costs per plasmid. We generally create kits...just the tool you need. The plasmids in this portion of the BIOFAB Plasmid Set contain terminators of ...populations. This kit consists of a 48 plasmid adapter library, with each plasmid carrying a unique barcoded Illumina-M13F...researcher at a nonprofit institution, you can order plasmids covering a wide range of fields from Addgene. ...kits to allow you to easily use a set of plasmids together on a tight budget. To help popularize our ...order from the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit... production (a variety of pExpression vectors) plasmids. The type IIS restriction sites used in these ...

... Read our Plasmids 101 blog series Find other plasmid protocols and tips Browse our plasmid cloning blog...alkaline lysis plasmid miniprep Starting material: 2 mL bacterial culture Product: plasmid DNA This kit-...kit-free plasmid miniprep protocol from Addgene follows a similar workflow as a column-based plasmid extraction... or agarose gel slices Product: plasmids or DNA fragments The plasmid purification and DNA gel extraction...depending on the type of DNA you’re purifying (plasmid, genomic, or DNA fragments from agarose).    In...causes the proteins and genomic DNA to precipitate. Plasmid DNA is free in solution. Proteins and genomic DNA... removed by centrifugation. At this point, the plasmid DNA sample can be treated with RNase to remove ...

...packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid which provides...AAV plasmid standard and AAV transfer plasmids. It’s easy to contaminate a workspace with plasmid, not...going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs which are...AAV’s protein shell, or capsid; 2) the pHelper plasmid which expresses adenovirus genes which help mediate...mediate AAV viral replication; and 3) the transfer plasmid which encodes a transgene of interest that’s packaged...structures that are prone to deletion from the transfer plasmid. The good news is there’s a few simple ways to ...multiple bacterial cultures of your AAV transfer plasmid at 30 °C instead of 37 °C and then screen for ITR...

...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...Many CRISPR plasmids are designed for lentiviral use, including Addgene’s top requested plasmid in 2015: ...itself due to a few missing proteins. The envelope plasmid contained a viral envelope to dictate the tropism...the vector could infect. Finally, the transfer plasmid encoded the desired transgene flanked by HIV-1 ...genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing lentiviral...recombination between the transfer and packaging plasmids does exist. To further reduce this possibility...

...encoded on separate plasmids and are co-transfected into cells alongside a plasmid encoding the minigenome...four viral genes in the same plasmid (as opposed to four separate plasmids), there are a few reasons arguing...separate (and thus smaller) plasmids. Finally, having four separate plasmids allows researchers to partially... 2 μg minigenome plasmid per well (in a 6 well plate format). However, since plasmid quality and transfection...Minigenome System). From a more technical standpoint, a plasmid with all four genes and necessary control elements...relatively large. Cloning the viral genes in this plasmid would be more difficult compared to if each gene...Technical tips It is important that all of these plasmids (those containing NP, VP35, VP30, L, and the minigenome...

...don't exactly "eat" plasmids as depicted in this comic, when a bacterium uptakes a plasmid, the genes on the... the plasmid can give them different phenotypes. For example, this plasmid encodes a green fluorescent... Read all of our Plasmids 101 blog posts Check out all of our blog posts on plasmid cloning Resources...comes from adjacent lysed bacteria and can include plasmid DNA or fragmented DNA released into the environment...membrane so the bacteria can easily uptake your plasmid of interest. Scientists can also use electroporation...exist as transient extrachromosomal DNA, like a plasmid, or it can integrate into the host bacterium’s ...cloning vector that contains both bacteriophage and plasmid properties. The phagemids are packaged into replication-incompetent...

... Hot Plasmid list. Other popular plasmids & blog posts @Addgene: The 10 Most Distributed Plasmid Technologies...inducible inactivation. Find the Robinson plasmids or the Royle plasmids and get to work! Robinson et al...designed the pOSIP (one-step integration plasmid) series of plasmids, vectors that convey both the sequence...mGRASP plasmids. Additionally, the lab recently described the use of another set of mGRASP plasmids in their... favorite plasmid technologies? Email us at [email protected]  and let us know what plasmids you would ...sgRNA libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA...new CRISPR screening tools, see our CRISPR/Cas Plasmids: Pooled Libraries webpage or read our blog post...

...Neurosci 2016. PubMed PMID: 26950006 Plasmid ID Plasmid Name Plasmid Type 74309 pcDNA3.0-TRPV4-p2A-ferritin-p2A-mCherry...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...features and applications of a partiular plasmid or set of plasmids. We hope that these articles make it ... you to find and use the plasmids you need. You can find all the hot plasmids from 2016 below. With over...over 45,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...stocks in a single plate with every plasmid or as individual plasmids shipped as agar stabs. Iverson, et...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...AAV-deliverable ABE.  Find the plasmid here Jesse Zalatan’s lab deposited plasmids for a chemically inducible...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...mGold, a new yellow fluorescent protein New CRISPR plasmids New items from the viral service A fast blue-light...protein.    Cells expressing an RFP reporter plasmid for recombinase activity were exposed to blue light...

...feature some recently deposited plasmids every quarter. The “hot plasmids” from last March were quite popular...behalf of scientists. 9. Plasmids 101: Positive and Negative Selection for Plasmid Cloning by Jennifer Tsang...past year: CRISPR, optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These...s the top 10 blog posts from the year. 10. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent... about mNeonGreen, mTFP1, and mWasabi and find plasmids containing these fluorescent proteins at Addgene..., and all possible base-to-base changes.   1. Plasmids 101: Transformation, Transduction, Bacterial Conjugation... Read other Addgene news Find the most recent plasmid blog posts Find the most recent CRISPR blog posts...

...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids Topics: CRISPR, Other CRISPR... where we highlight a few of the newest CRISPR plasmids available at Addgene. If you want to read more...will cover: Miniature CRISPR-Cas systems SEND plasmids Prime editing improvements Miniature CRISPR-Cas...Read the paper in Molecular Cell Find the CasMINI plasmids Efficient CRISPR editing with a hypercompact ...paper in Nature Biotechnology Find the Cas12f-GE plasmids Programmed genome editing by a miniature CRISPR-Cas12f...cells using various delivery methods, including plasmid, ribonucleoprotein, and AAV.  Read the paper in... in Nature Chemical Biology Find the AsCas12f1 plasmids   Selective endogenous encapsidation for cellular...

... requested plasmids. I wondered how many scientists have used the protocol for this plasmid, and if it... your protocols onto your plasmid page, scientists who request your plasmid will have easy access to your...restriction digest of plasmid DNA, and bacterial transformation. And more plasmid-related educational resources...be found in our Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene? Check... resources, such as protocols, Plasmids 101 articles on various plasmid features, and reference tables...deposit their plasmids to the repository. These protocols show up on the related plasmid pages and can...http://www.addgene.org/ If you’re working with plasmids and you decide to share them using Addgene’s repository...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...write about a recent plasmid deposit please sign up here.  Listen to the hot plasmids episode! New, highly...hundred SH3 domain containing plasmids available through Addgene. These plasmids can be used to carry out ...dsRNA necessary for RNAi. The Vellai lab’s new plasmid, T444T, contains T7 terminators adjacent to the...to increased RNAi efficiency. Find the T444T plasmid here! Strum A, et al. Nucleic Acids Research. 2018... Drosophila and C. elegans.   Find the TurboID plasmids here! Branon et al. Nat Biotechnol. 2018 PubMed...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... recent plasmid deposit please sign up here.      Listen to this episode of the Hot Plasmids podcast ... (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These ARG plasmids contain...responses in C. elegans. Find the split BoTN/B plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed...potentially problematic for imaging. Find the ARG plasmids at Addgene. For more details on acoustic reporters...s collection consists of a library of reporter plasmids, each containing two fluorescent proteins and ...