...Addgene Blog: Plasmids 101: How to Verify Your Plasmid Plasmids 101: An Inside Look at NGS Plasmid Quality ...results of the plasmids in our repository with known reference sequences, such as full plasmid sequences provided...up their plasmid and cloning verification steps and free up more time for using your plasmids instead!...Snapgene-powered plasmid maps. Resources on Addgene.org Sequence Analysis of your Addgene Plasmid Addgene Plasmid...provided by the laboratories that deposit their plasmids with us or other entries in NCBI’s numerous databases... grown over the years (we now have over 60,000 plasmids!), the number of sequencing results we analyze...busy week, we may need to analyze more than 200 plasmids as part of our quality control process. Consequently...

... target plasmid and pX330-sgRNA plasmid (Mashiko et al. 2013). The pCAG-EGxxFP target plasmid contains...pCAG-EGXXFP plasmid along with the individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA... (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences ... system floating around, and many pre-designed plasmids containing these variants ready for purchase. ...multi-cloning site (MCS) between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and... you can insert this region into a pCAG-EGXXFP plasmid using standard cloning techniques (The XX indicates...the EGFP fluorescent signal, for details on this plasmid, see Mashiko et al., 2013). Testing the efficacy...

...your expression plasmid from these strains is not recommended as these auxillary plasmids may be isolated...control such as the pLys plasmid, which suppresses basal T7 expression. The pLys plasmid contains a chloramphenicol...arabinose-inducible plasmid will not express in an IPTG induction strain for example, nor will a p15 plasmid be compatible... for More Plasmids 101 Articles?  Find them all here: http://blog.addgene.org/topic/plasmids-101   ...In a previous Plasmids 101 blog, we reviewed the salient features of several popular strains of E. coli...strain? Protein expression from high-copy number plasmids and powerful promoters will greatly exceed that...BLR (DE3) Tetracycline RecA-deficient; best for plasmids with repetative sequences.  F- ompT lon hsdSB...

...example (the top requested plasmids from 2015 and 2016 were CRISPR plasmids), but so are lentiviral vectors...reason, the top requested plasmid of 2017 is the lentivirus envelope plasmid pMD2.G from Didier Trono’...Plasmid technologies are constantly evolving, but sometimes a technology is so useful it forever enhances...’s lab! Lentivirus envelope vector pMD2.G This plasmid has been requested nearly 6,000 times (over 800...the broad usefulness of pMD2.G and VSV-G. This plasmid has been used in studies of the secretome, ubiquitin...to check out the Trono Lab website. If you have plasmids you’d like to deposit with Addgene, be sure to...Who knows, maybe yours will be the top requested plasmid of 2018! Additional Resources Quick Guide to ...

... awarded plasmids from Addgene as a successful applicant to the Addgene, Seeding Labs Plasmid Grant. For...Learn about the Addgene, Seeding Labs Plasmid Grant Read about Plasmid Grant Recipient, Gladys Kaba Resources... fellow nonprofit, Seeding Labs, to distribute plasmids to researchers in developing regions and thereby...accelerate their research. Kwabena came across the plasmid grant in Seeding Lab’s Newsletter and recently ...plans to use his research experiences as well as plasmids from Addgene to bring new opportunities to Ghanaian... Kwabena applied for the Addgene, Seeding Labs plasmid grant. 7:12 - 7:11 : Kwabena's work in Christopher... recent recipient of the Addgene, Seeding Labs Plasmid grant and is working on a variety of projects in...

... As reviewed in our Plasmids 101 series, proteins are overexpressed from a plasmid construct, most often...multiplexing. pCri plasmids are divided into two sets, each of which includes plasmids with different varieties...on the Addgene Blog Plasmids 101: E. coli strains for protein expression Plasmids 101: Protein tags ...varieties of tags, both N- and C-terminal. Each set of plasmids uses the same restriction enzymes for cloning ...for one protein quickly and easily. These tagged plasmids also include cleavage sites should one need to...other tags used in the pCri system, please see Plasmids 101: Protein Tags. Beyond E. coli: purifying ...subtilis and Pichia pastoris pCri also includes plasmids designed for protein expression in systems other...

... circularized plasmids from plexWell™ NGS data. Once Addgene produces plates of plasmid DNA, they are ...blogger Joe Mellor, Founder and CEO of seqWell Inc. Plasmids and PCR products are the bread and butter of molecular...using these methods for routine processes like plasmid validation. At seqWell, Inc., our mission is to...Addgene’s large and growing collection of curated plasmids from all over the world. The rest of this piece...partnership with Addgene to sequence large numbers of plasmids. plexWell™ technology overview For any NGS experiment...individual input DNA samples (e.g. a collection of 96 plasmids on a plate) will typically have a wide range of...to, for example, assemble hundreds of different plasmids from a single Illumina MiSeq® run. As shown in...

...Chemogenetics Plasmid Serotype Name 119472  AAV5  AV SYN PSAM4 GlyR IRES EGFP   Biosensors Plasmid Serotype...inventory of serotype testing AAV New control AAVs Plasmid Serotype Name 105530  AAV1 pAAV.CMV.PI.EGFP.WPRE.bGH...biosensor AAV inventory New glutamate sensors Plasmid Serotype Name 106176  AAV1  pAAV.hSynapsin.SF-iGluSnFR.S72A...pAAV.hSynapsin.SF-iGluSnFR.S72A New calcium sensors Plasmid Serotype Name 104496  AAV1 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE...pAAV-hSynapsin1-FLEx-axon-GCaMP6s New voltage sensors Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST...of DREADDs / Chemogenetics AAVs New DREADDs Plasmid Serotype Name 75033  AAV1 pAAV CD68-hM4D(Gi)-...pAAV SYN1 HA-hM3D(Gq) New chemogenetics AAV Plasmid Serotype Name 119741  AAV5  AAV SYN flex PSAM4...

...service features 210 plasmids: empty backbones, fluorescent protein tags, SARS-CoV-2 plasmids, etc. It is our...to Elute and Store Your Plasmid DNA? by Paolo Colombi Should I store my plasmid in TE, Tris, or water?...Here we cover five methods to do this. 5. Plasmids 101: Plasmid Incompatibility by Leah Schiesow If you're... We've had 11 blog posts on COVID-19 featuring plasmid tools from depositors who quickly shared their ...you're thinking about hosting multiple plasmids in one cell, it might be that you can't stably maintain...This blog post features two mechanisms that cause plasmids to be incompatible. 4. RNA Extraction Without...CRISPR to detect SARS-CoV-2, including many with plasmids deposited at Addgene! 2. DNA Purification Without...

... in one place. Plasmid mapping These tools are for viewing, editing or making plasmid maps, but can also...make plasmid maps. Free for academic users. Serial Cloner: Free desktop-based software for plasmid design...design and mapping. ApE (A plasmid Editor): A free, donation-based plasmid analysis tool including editing...Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid pET21a-BirA.   DNA / ...Primer design. Plasmid mapping. DNA sequence analysis. We all have our favorite tools for tackling these...The free SnapGene Viewer is great for looking at plasmid maps and viewing sequencing traces, while the paid... paid version provides more tools for plasmid mapping and design (Figure 1). Benchling: While you might...

...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...

...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...

...modifications Learn more molecular biology in our Plasmids 101 eBook! Researchers use various techniques ...for your protein of interest is inserted into a plasmid expression vector that is then transformed into...being able to help the scientific community share plasmids.   Additional Resources Thermofisher Protein ...Addgene.org Bacterial expression systems Search for plasmids by expression system ...

...Resources on the Addgene Blog: Plasmids 101: The Promoter Region Plasmids 101: Cre-lox Light-Switchable...Promoter Subtype Promoter Example Activator Addgene Plasmid/Kit Alcohol-regulated AlcA promoter AlcR pGGA008...use light to regulate transcription. Red flame plasmid pDawn contains the blue-light sensing protein YFI...Systems for E. coli Browse other blog posts from our Plasmids 101 series Resources on Addgene.org: Inducible...Expression Systems at Addgene Cre-lox Guide and Plasmids     ...

...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...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...head over to our CRISPR Plasmids and Resources page.  CasPhi is 70 kDa Cas protein identified exclusively...offers advantages for cellular delivery. Get the plasmids at Addgene Sc++ is a codon-optimized, engineered...nuclease that has a broad 5′-NNG-3′ PAM. Get the plasmids at Addgene The Human GlycoGene CRISPR Library...

...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...need. If you'd ever like to write about a recent plasmid deposit please sign up here. Here's what you'...easily be applied to other systems.    Find BLITZ Plasmids here! Xiong, et al. eLife. 2021. https://doi.org...from Goertsen D, et al., 2022.  Find M-CREATE plasmids here!  Goertsen D, et al. Nature Neuroscience....have higher risks of off-target mutations. These plasmids contain catalytically inactive dCas9 and microbial...Image from Wang, et al, 2022.  Find dCas9-SSAP plasmids here! Wang et al. Nat Cell Biol. 2022. https:/...

...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...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'...Ladder  by: Rachel Leeson This simple, inexpensive plasmid-based ladder system is a cost-effective alternative... ladders. Image from Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60...kD proteins or 20, 40, 60, and 80 kD proteins. Plasmids containing 150 or 250 kD proteins are also available...

...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. .... Today, it's time for another edition of Hot Plasmids!  Here's what you'll find in this post:  New...., 2023 under CC-BY 4.0 License. Find PE6 plasmids here!  Doman, J.L., Pandey, S., et al. (2023)....therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D., et al. (2023). Cell type-specific...pulse-chase experiments, and more. Find LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID...LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments open up new possibilities... promoter activity in single cells. Find iPAK4 plasmids here! Lin D., Li X., et al. (2023). Time-tagged...

...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...