...blog Addgene's Tips for Plasmid Quality Control Plasmids 101: In-house NGS Plasmid Quality Control Now at...sequencing all incoming plasmids. If you are interested in requesting one of these plasmids, you can request ...data for each of their plasmids, from descriptions of key components of the plasmids to partial, full, or...to ensure all plasmids pass our high-quality standards and contain the expected plasmid features.  ...Once Addgene receives deposited plasmids, they undergo complete plasmid sequencing using an in-house Illumina...will compare the NGS results to the plasmid information, other plasmids in the deposit, NCBI sequence and...results and deposited sequences and plasmid information.  For each plasmid deposited at Addgene, QC scientists...

... best plasmid for your research can be a daunting challenge. With over 80,000 deposited plasmids from ...through to the plasmid search results: More than 200 different GCaMP containing plasmids are available...search tool lets you access plasmids from journal articles. From paper to plasmid Authors regularly indicate... to include specific plasmid ID numbers. If you know the ID number for a plasmid, your straightforward...page to see the plasmids deposited by our own CSO. But what if you don’t have plasmid ID numbers, or you... Not every plasmid search will start from a publication. You may have an idea for a plasmid that would...exact plasmid you need. The ability to apply a wide range of filters isn’t new to Addgene’s plasmid search...

...deposits with more than 10 plasmids is to use our deposit spreadsheet?  Our plasmid deposit process can be... Send plasmid information to Addgene Material Transfer Agreement (MTA) approval  Send plasmids to Addgene...spreadsheet and copy and paste your plasmid data directly into our file, one plasmid per row, as outlined by the... Data Field Description Plasmid Name Use a descriptive plasmid name and match the name as it ...application of the plasmid and will help a scientist outside your field understand if this plasmid will work for...possible, we propagate plasmids in the standard cloning strain DH5α. For plasmids with highly repetitive...lentiviral, retroviral, and AAV plasmids, choose the NEB Stable strain. For plasmids containing the ccdB gene...

...curating plasmid collections. Addgene's "How to cite this plasmid" section Anytime you use a plasmid from ...process, we explain how to cite plasmids from Addgene right on our plasmid pages and provide a citation ...comparing two plasmids, we recommend reviewing the cited papers to determine whether the plasmid has been ...collection or if you think a particular plasmid or type of plasmid is missing from our collections pages...spotlight some of the plasmids that have come through our doors in our “Hot Plasmids” section. We’ve also... added the Plasmid of the Day feature to our home page - each day we showcase a new plasmid that you might...variety of plasmids in the repository. And last, but certainly not least, we often feature new plasmid technologies...

...What SARS-CoV-2 plasmids from Ginkgo Bioworks are available for request? Currently, plasmids containing all...available. Plasmids are continually added as they complete QC and you can find all available plasmids listed...to fresh data! These same plasmids are then added into our normal full plasmid NGS queue in order to bring...pipeline. For the first ~50 plasmids we perform our standard full plasmid sequencing by NGS. Addgene’s...on Addgene.org Find all plasmids from Ginkgo Bioworks  Find COVID-19 plasmids for your research Browse... to synthesize and share over 1,850 SARS-CoV-2 plasmids to help academic and industrial labs accelerate...confirmed the expected features and sequences in each plasmid, none of these constructs have been functionally...

...materials that we use in our labs, including plasmids. All plasmids at Addgene have unique ID numbers associated...identify a plasmid available through Addgene and find verified information about each plasmid item. Given...associated plasmid page on the Addgene website and find specific information about the plasmid: name, description...description, how the plasmid was generated, validated sequence information, and more. The plasmid page also helps...publications that have used the same plasmid through the “Articles Citing this Plasmid” section. Having IDs linked...specific plasmids used. RRIDs and resolvable compact identifiers now exist for all Addgene plasmids! In an...our example plasmid, lenti-Cas9-VQR-Blast. If you scroll down to the bottom of this plasmid’s page you will...

...viral vectors and plasmids with repeating sequences. Repeating sequences within a plasmid can undergo intramolecular...contain the full length plasmid.   Figure 1: The full lentiviral plasmid is expected to be >12 kb...in my samples? Plasmid recombination can affect viral vectors (LTRs and ITRs), plasmids with multiple ...other plasmids that contain repeating elements. Knowing how to amplify recombination-prone plasmids and ... length plasmid. If you find that your bacterial cultures contain a mix of full length plasmid and the...smaller, recombined plasmid around 1.5 kb. When cut with BamHI, the expected linear plasmid is present, but...get to see everything that can go wrong with a plasmid. One of the most frequent problems that I help ...

...protocols, and, of course, plasmids. Today we’re trying a new way to present plasmid info with a new segment...The Hot Plasmids Segment. Click on the player below to listen to a quick (~5 min) Hot Plasmids podcast... new plasmid technologies from one of our recent newsletters. Listen to the entire Hot Plasmids episode...greatest plasmids through this podcast segment. Below, we break down each of the Hot Plasmids segments...articles on the Hot Plasmid webpage. We hope you enjoy this new way to learn about plasmid technologies. If...comments about the Hot Plasmids Podcast Segment, or have recommendations for new Hot Plasmids topics, let us ... comments section below. Hot plasmids covered in this segment Hot Plasmid 1: CRISPR X from Michael Bassik's...

...and an envelope expressing plasmid, such as our second most requested plasmid of all time, pMD2.G. Find...wide range of applications. This plasmid was the top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB... top requested plasmid from 2016. You may notice the “V2.0” at the end of this plasmid which indicates...prior version of this plasmid. Indeed the first version is the 15th most popular plasmid of all time, but ...envelope expressing plasmid pMD2.G. Find pMDLg/pRPE. scramble shRNA - This plasmid from David Sabatini....1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...15 years of plasmid sharing has certainly taken us on many adventures...From moving office locations ...

... here at Addgene – 500,000 plasmids shipped! That's a 1/2 million plasmids sent to happy scientists around... you all the plasmids you need. We're focused on our mission – documenting your plasmids, storing them...500,000 plasmids go? Far and wide – that's where! Check out the map below which depicts where plasmids have...Sidney Street. Then the plasmids travel to labs around the world. We've shipped plasmids to more than 90,000...providing quality control and customer support, making plasmids easy for you to find and request in our online...shipping them to you quickly.   So, what does plasmid shipping look like at Addgene? Here's an inside... workings in Addgene's lab. We ship individual plasmids as transformed bacteria in stab culture format...

... Requested Plasmid in 2015 Find Hot Plasmid Technologies Write about Your Favorite Plasmid Technology ... indicates that a plasmid has been requested 20+ times   A red flame means a plasmid has been requested...answering plasmid requests can be a time-consuming chore for labs that have popular plasmids but lack ...All plasmids deposited at Addgene enhance scientific sharing worldwide but a select few are particularly...particularly special. Over 1000 of our 60,000+ deposited plasmids have been requested more than 100 times each, ...to sharing epitomized by the creators of these plasmids with the first annual Blue Flame Awards. What'...scientists around the world choose to share their plasmids through our repository since we know biological...

... we use SnapGene for plasmid quality control are: To identify important plasmid features To align NGS...design experiments for further plasmid verification Identifying plasmid features While our quality control...control process for incoming plasmids can vary based on the contents of a plasmid, the first step is always...send the plasmid off for Sanger sequencing.     If a repetitive region is present in the plasmid, we might...sequencing results for plasmids in the repository at a glance. Our Snapgene-powered plasmid maps and sequences...the Addgene Blog How to verify your plasmid Check out our Plasmids 101 featured topic page Additioinal...results to perform quality control on deposited plasmids. We’ve also talked about how our new Snapgene ...

...mutagenesis of Plasmids. A) Generation of mutagenized plasmids. PCR primers (green) amplify the plasmid template...with the nuclease-resistant nicked plasmid (the PCR product). Plasmids are isolated from the resulting colonies...the final circular plasmid product pictured here. B) Screening of recovered plasmids for the mutation of...Smaller plasmids (~3 kb) are generally more efficiently amplified than larger constructs, but plasmids as ...DpnI) to eliminate the parent plasmid from the PCR products, the plasmid template should be isolated from...have taken up the mutagenized plasmid. Note that any residual parental plasmid (usually from incomplete DpnI...additional restriction site into your plasmid, then, when the plasmid is digested with the appropriate restriction...

...cells to retain plasmids - if a cell does not inherit plasmid due to improper plasmid partitioning, it...unstabilized plasmids were retained in less than 10% of cells while plasmids containing the plasmid addiction...ensure that plasmids are passed down through generations (Figure 1). These so called “plasmid addiction ...only survive if they retain the plasmid. Since their discovery, plasmid addiction systems have been used...found that if a mini-F plasmid carried a specific 700 bp segment of DNA, the plasmid was propagated better...selection in one plasmid, they were able to create an efficient cloning tool. The plasmid pKSCTMF contains...restriction-modification systems onto plasmids allowed cell populations to retain more of these plasmids as compared to those...

... (A) Cumulative CRISPR plasmids shipped by year. (B) Cumulative CRISPR plasmids deposited with Addgene...,000 CRISPR plasmids in our repository, and we have shared almost 300,000 CRISPR plasmids and 12,000 CRISPR...these CRISPR plasmids (and libraries)? We wanted to know, so we classified CRISPR plasmids containing a...CRISPR plasmids requested each year by year of deposit (Figure 3), we saw that once CRISPR plasmids become... CRISPR plasmids seem to never truly go out of style. Instead, an almost even mix of plasmids (by age)...Total CRISPR plasmids deposited by country; square size is proportional to number of plasmids deposited ...Total CRISPR plasmids requested by country; square size is proportional to the number of plasmids requested...

...cost required to make a plasmid. In 2023 (and at closer to 2 million plasmids shared than not!), we think...great Plasmid Debate of ’18, Blue Flame depositor John Doench calculated the time to make a plasmid. Doench... make a plasmid? Doench also calculated the cost of consumable reagents used to make a plasmid, which ... to make plasmids that already exist, and we significantly reduce costs of producing plasmids with high-throughput...lot of plasmids, it’s almost always faster to order from us!) Ask the PI If you want a plasmid for free...high-throughput” Instead of making one plasmid at a time, try constructing several plasmids in parallel. That way, you...collection pages to more easily find plasmids Search for a plasmid in Addgene’s repository ...

...cloning vector. Lentiviral Plasmids -  If you're familiar with the pLKO plasmids you'll note that those are...Actually, these two plasmids from Didier Trono's lab are our two most popular plasmids of the past 10 years...influx of plasmids for reprogramming differentiated cells into pluripotent stem cells. Plasmids from Shinya... have 19 different ubiquitin plasmids in the repository. 100,000 Plasmids Shipped! - By 2010, Addgene ...our stride. We shipped our 100,000th plasmid. A variety of plasmids continued to be popular years after...with all the new plasmids. So far, we have distributed over 10,000 CRISPR/Cas9 plasmids to more than 3,...10 years of plasmid sharing  Interested in more facts about Addgene's 10 years of plasmid sharing? Check...

...multiple gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the...rectangles) are cloned into various plasmids using oligonucleotides. These plasmids contain Type IIS restriction...different purposes.   Gersbach Lab multiplexing plasmids: This plasmid set allows you to express 2-4 gRNAs, with...assembled to produce a plasmid encoding the gRNAs and Cas9. As with the Gersbach lab plasmids, multiple Cas9 ...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids allow you to assemble 2-4 gRNAs through...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids can be used to express up to 8 gRNAs after...Yang Lab Single Transcript Multiplexing Plasmids: These plasmids are similar to the Csy4 polycistronic ...

...replication. In general, these plasmids tend to be low copy number. Conversely, plasmids that can initiate DNA ...growth against sustainability. Plasmid DNA replication and regulation Plasmids must also strike a balance:...a given plasmid has been reached, negative feedback circuits reduce replication of the plasmid to balance...balance the total plasmid number.  Let’s consider pSC101. This plasmid is stringently controlled and has...promotes initiation at the plasmid ori, after which replication of the plasmid proceeds (and terminates)...tool to stringently regulate plasmid replication. For example, the plasmid ColIb-P9 encodes the protein...All plasmids rely on their host cell's replication machinery in order to replicate—but not always to ...

...consider your plasmid's compatibility with one or more other plasmids. Generally speaking, plasmids with the...enabling a plasmid to reproduce itself as it must to survive within cells. The replicons of plasmids are generally...cell. Figure 1: A plasmid map showing the standard features of a plasmid. So, how do you choose...have only one plasmid type in your cells at a time? Do you want to make a lot of your plasmid? Is the gene...avoid plasmid incompatibility In other words, the best choice of ori depends on how many plasmid copies...a rule, plasmids from the same group should not be co-transformed, so if you need two plasmids for an ...purify your plasmid DNA: The insert Bacteria tend to maintain fewer copies of plasmids if they contain...