...loved helping to answer your plasmid and cloning questions through our Plasmids 101 series and by responding...Melina Fan Plasmids 101: The Promoter Region – Let's Go! By Kendall Morgan Plasmids 101: Mammalian...Joanne Kamens Plasmids 101: Antibiotic Resistance Genes By Marcy Patrick Plasmids 101: Yeast Vectors...FAQs Answered! By Caroline LaManna Plasmids 101: What is a plasmid? By Margo R. Monroe Best wishes...as seen through the lens of Addgene’s blog and plasmid repository. Our blog is just over a year old, and...detailed information about this new technology. Our Plasmids 101 series continues to grab scientists’ attention...Genome Editing By John Doench (Guest Blogger) Plasmids 101: Origin of Replication By Kendall Morgan ...

... multiple plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually...encodes VSV-G, a packaging plasmid containing structural genes and a transfer plasmid carrying the transfer...Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...and packaging plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection...By dividing the viral elements across multiple plasmids, the risk of recombination and consequently, the...extra level of safety, they also require a fourth plasmid. The requirement for additional DNA can impact ...thereby affect the titer of the virus. Additional plasmids will also affect experiment optimization. Before...

...about Popular Plasmids in the Repository Learn about New Plasmid Technologies in our Hot Plasmid Articles ...to create a plasmid based on the biotin ligase BirA backbone (Eric Campeau, Addgene plasmid #26624) was...inserting BirA into a pMal plasmid (NEB) thus creating the pMal-T-Avi-His/BirA plasmid now available through...other Hot Plasmids Find Additional Posts on Mol Bio Techniques Find Additional Posts on Plasmid Technologies...controlled easier than formation of GST dimers. Avi-Tag plasmid for improved biotinylation selectivity A common...exploit the best of both worlds. We generated a plasmid that produces biotin ligase and a protein of interest...and a 6X HisTag for affinity purification (see plasmid map and cartoon below). Including biotin in your...

...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 Podcast!  MarathonRT...’s lab at Yale University recently deposited a plasmid for expressing a novel reverse transcriptase (RT...polymerase processivity. Please find the MarathonRT plasmid here! Zhao C, et al. RNA. 2018 PubMed PMID:29109157... cell surface. Their toolkit includes a set of plasmids each containing an anti-GFP nanobody fused with...fluorophores. Find all of the nanobody toolkit plasmids here. Buser DP, et. al. PNAS 2018. PubMed PMID...

... presence of a plasmid is disadvantageous from the bacterium's perspective – a plasmid-containing cell...to maintain the plasmid. Adding an antibiotic resistance gene to the plasmid solves both problems at once...Biology Reference Pages View our Plasmid Protocols Find Your Perfect Plasmid Backbone ...think about how much easier it makes our lives. Plasmid transformation into E. coli is a fairly inefficient... which cells successfully received the correct plasmid, scientists would spend hours to days trying find...cell must replicate the plasmid in addition to its own chromosomal DNA, costing additional resources to ...once – it allows a scientist to easily detect plasmid-containing bacteria when the cells are grown on ...

...Collection page, you can browse for plasmids by species and find our guides, plasmid kits and other resources for...microbial research. Kits, guides, plasmids, and more We have Plasmid Kits for microbial work with fluorescent... To make it easier for you to find plasmids for your work, plasmids in the Microbiology Collection are... a useful plasmid that’s not in the collection? Tell us about it with our Suggest a Plasmid Form! Finally...us at [email protected]. If you are looking for plasmids used in E. coli and yeast, you’re in luck! Addgene...develop resources for your community! To contribute plasmids made in your lab, see www.addgene.org/deposit ...

...mutation Improves plasmid yield F Host does (F') or does not (F-) contain the fertility plasmid. A low copy-number...and general DNA repair Reduces plasmid recombination, increases plasmid stability recBCD Exonuclease...Vectors Looking for More Plasmids 101 Articles?  Find them all at our Plasmids 101 Topic Page!     ...Nov 14, 2017. You've worked hard designing your plasmid – you carefully selected the optimal promoter for...labor-intensive process of replicating your new plasmid: the Escherichia coli bacteria. It’s hard to count...whether it is appropriate for propogating your plasmid or carrying out your experiment. History of E. ...major improvements such as those that increase plasmid yield and/or DNA quality. Table 1 below outlines...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...paper in Nature Communications Find the Cas12b plasmids  Plant base editing with a Cas9-adenosine deaminase...wheat. Read the paper in Genome Biology Find plasmids containing these base editors Spatially and ...Neuroscience Find the AAV-based SaCas9 genome editing plasmids String assembly gRNA cloning (STAgR) String ...the paper in PLoS One Find the STAgR cloning plasmids Mobile-CRISPRi allows CRISPRi experiments in...Gross’s lab recently deposited “Mobile-CRISPRi” plasmids, a set of modular CRISPRi tools that can be used...

... Can I use my favorite plasmid with this protocol? Maybe. Not all plasmids will work for the one-pot...strands of the plasmid will be nicked during the first BbvCI digest and the whole plasmid will be degraded...those in the plasmid. Again, if they are in the opposite direction, both strands of the plasmid will be nicked...map of your plasmid. If the sites are in opposite directions, you just need to use a plasmid with BbvCI...Mutagenesis to Mutate Your Plasmids Resources on Addgene.org Find Plasmids from the Whitehead Lab Use.... This technique relies on the generation of a plasmid library that expresses all desired variants of ... a selective pressure winnows the pool down to plasmids expressing variants with optimal function. High-throughput...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...Read the paper in Science Find the new SHERLOCK plasmids More precision with dual-nuclease Cas9-Cas9 ...Nature Communications Find the Cas9-Cas9 chimera plasmids A modular toolkit for genome editing in plants...post Read the paper in Science Find the Cas14 plasmids Anti-CRISPR proteins for optogenetic control...paper in Nature Methods Find the anti-CRISPR plasmids Tagging human transcription factors with GFP...White’s lab recently deposited a set of CRISPR plasmids to tag human transcription factors with GFP. As...

...experiments in planarians with their recent plasmid deposit. These plasmids enable two main types of experiments... cloned into plasmids to investigate for each of these planarian species. These plasmids all have a PR-T4...insert the cDNA into the desired plasmid backbone. The Sánchez lab plasmids allow students and new labs to...Resources at Addgene.org Find plasmids from the Sánchez lab Find plasmids for other developmental models...of specific gene expression. The 49 deposited plasmids enable students to study gene expression and functions... dsRNA-induced RNAi experiments. Notably, this plasmid harbors a multi-cloning site flanked by T7 promoters... produce DNA templates via PCR off of the same plasmid and use these to create riboprobes for in situ ...

...γ-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G (Vesicular...in trans by specific plasmids. This method, therefore, reduces the number of plasmids that are required ...encoded by the transfer plasmid, but instead are provided in trans by other plasmids or the packaging cell...do many γ-retroviral plasmids have similar names? The nomenclature of the plasmid backbone is meant to...components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA of interest..., which facilitate integration of the transfer plasmid sequences into the host genome. Typically, it is... genome upon viral transduction. Most transfer plasmids available from Addgene were derived from MoMLV...

...specifically, the Addgene-Seeding Labs plasmid grant provide plasmids to these researchers. In the third ...the Addgene-Seeding Labs Plasmid Grant Check Out Addgene-Seeding Labs Plasmid Grant Awardees in 2016 and...to provide scientific resources, equipment, and plasmids to researchers in resource-limited settings. More...third round of the Addgene-Seeding Labs plasmid grant we are excited to introduce the 5 awardees and their...biofilms formed on various structured surfaces. Plasmids expressing fluorescent proteins will be used to...microscopy and FACS. In an additional approach, plasmids such as pACBSR-hyg and pFLP-hyg will be used to...regulation and diabetes will be expressed from plasmids (find more information on bacterial expression...

...organizations choose to share their plasmids under an OpenMTA, which allows plasmids to be used for both research...If you’ve deposited or requested plasmids from Addgene, then you’ve been involved in the material transfer...Even though this process is essential to sharing plasmids around the globe, it can be a bit of a mystery...use agreements. The UBMTA, which is the MTA most plasmids at Addgene are shared under, is well-understood...we are not a party (signatory) to the MTAs for plasmids in our repository. Instead, we facilitate the ...institutes. But they’re not! The vast majority of our plasmids are distributed under the UBMTA, which is widely... our process that differs.  When you deposit a plasmid with Addgene, your organization’s tech transfer...

...beneficial for plasmid clearance. It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only.... pCutamp cures the plasmids by targeting the ampR promoter in all three plasmids. pCutamp is then removed... 13 plasmids with 14 rounds of transformation in 30 days. Figure 2: (A) MUCICAT plasmids for ... original pCas/pTargetF system consists of one plasmid that expresses the S. pyogenes Cas9, a temperature-sensitive...the pTargetF construct and then introduce both plasmids into the cell. The edit in the chromosome can ...the pMB1 replicon, resulting in cutting of the plasmid. At this point, subsequent introductions of pTargetF...addition of the sgRNA sequence in pEcgRNA. Next both plasmids are transformed in E. coli with the donor DNA ...

...resistance DNA plasmids are traditionally grown in bacteria as a way to produce more of the plasmid, isolate...isolate individual plasmids, or express a gene contained in a plasmid. During the propagation and/or transformation...receive a copy of the plasmid or will lose it during division. These non-plasmid-containing bacteria can...Over-Agar Antibiotic Plating Plasmids 101 eBook Resources on the Addgene blog Plasmids 101: Antibiotic Resistance...Resistance Plasmids 101: Blue-White Screening Choosing Your Perfect Plasmid Backbone  ...Plasmids need antibiotic resistance genes so that you can specifically isolate bacteria containing your...your experiment — to grow, express, or isolate plasmids. The presence of an antibiotic resistance gene...

... Addgene/Seeding Labs plasmid grant and plans to use the grant to order plasmids she can use as positive...similar grants back in June, Gladys will be using plasmids in creative ways that we wouldn’t necessarily ...HR HPV types, predominantly HPV 16 and 18.” HPV plasmids as positive controls To start her work, Gladys...cytological tests currently in use. Gladys will use plasmids containing a variety of HPV genes as positive ...incredibly happy to hear that Gladys will be using plasmids in a way that can potentially have a direct impact...awardees We will be opening the Addgene/Seeding Labs Plasmid Grant to a third round of applicants at the start...Seeding Labs for this continued opportunity to get plasmids to the researchers who need them the most. Check...

...methionine (i.e. the plasmid complements the deletion). Find the Plasmids Here Plasmid features enable multiple...the overexpression plasmids. Finally, students determine if the presence of the plasmid corrects any known...diverged. Tips on using the Pathways Over Time plasmids The plasmids are used as part of a semester-long research...less-well studied organism into overexpression plasmids. Students next transform deletion strains from...phenotypic abnormalities due to the deletion (i.e. the plasmid complements the deletion). If the phenotype is ...cerevisiae met deletion strains with overexpression plasmids carrying MET/Met genes from either S. pombe or.... Students then determine if expression of the plasmid-encoded MET gene allows the transformed strain ...

...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...

...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...