...This post is part of our ongoing Plasmids 101 series. Plasmids 101 will provide you with an overview ...of the Plasmids 101 posts here. Now that we have covered antibiotic selection here at Plasmids 101, we...α-complementation plasmid. This way you can quickly and easily identify colonies that not only have your plasmid (antibiotic...rescued by genes supplied on the transformed plasmid. Such plasmids may contain genes that allow for the use... able to select for colonies that contain your plasmid is a great start when cloning, but how about being...being able to choose those that contain plasmid with an insert? Blue-white selection is a widely used ...far, so good, but how does this help us screen plasmids? Blue-white screening in the lab Scientists discovered...

...Addgene Blog Plasmids 101: Common Lab E. coli Strains Plasmids 101: TOPO Cloning Plasmids 101: Golden ...-containing vectors in mind, any plasmid that contains the F plasmid (F’ strains) will also be resistant...... The ccdB gene, located on the F sex factor plasmid of E. coli, is part of a toxin-antitoxin system...encoded by the ccd operon, which is responsible for plasmid maintenance during cell division. ccdB codes for...Cells that lose ccdA through the loss of the F plasmid, succumb to the toxicity of CcdB.   Figure...disrupted allowing cells expressing the recombinant plasmid to propagate. Any cells that contained non-recombinant... of clones that do not contain the recombinant plasmid and therefore makes the cloning process much more...

...fragmenting plasmid DNA, PCR product, or chromosome. One of the main causes of degradation during plasmid DNA ... a kit Read our blog posts about plasmid cloning Browse our Plasmids 101 blog posts Resources on Addgene.org...Biology Reference Find protocols on plasmid cloning Find plasmids for your research ...Nowadays, the most common E. coli strains used for plasmid amplification are endA negative, meaning the gene...degradation and which experiments you can use your plasmid.  DNA elution choices: TE, Tris buffer, or water...polymerases. In the situation where you need to use plasmid in TE for any enzymatic reaction, an effective ...choices that companies offer for the elution of plasmid DNA through their kit. The advantage of using a...

...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...small collection of plasmids for coronavirus research, including a handful of plasmids that have been in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from... labs also deposited plasmids for producing COVID-19 diagnostic enzymes. Plasmids in this toolkit include...These plasmids contain both untagged and tagged versions of SARS-CoV-2 ORFs and include plasmids that ...scientists through plasmid sharing and more. As you’ve read above, some of these plasmids have been in the...

...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 Hot Plasmids! Optogenetic...Grefen and Blatt, 2012). The 2in1 Plasmid Toolkit consists of 4 plasmid toolkits generated with 4 different...deposited the resulting optogenetic plasmid tools which includes plasmids encoding a fusion between cPYP and...targeting and editing experiments. Find the CASANOVA plasmids here! Bubeck F et al., Nature Methods 2018. PubMed...as mice, C.elegans, and Zebrafish. Find Archon1 plasmids at Addgene! Piatkevich KD et al, Nat Chem Biol...

...restriction digest, and then plasmid sequencing if you can. To quote Doench, “Whole plasmid sequencing is just ...correctly banded clones for plasmid sequencing and found 98% of the plasmids were perfectly correct. Because... Mammalian Plasmid pRDA_791 (contains only orio and ampR) Drosophilia Plasmid pRDB_051 (for...the assembled plasmid. It shows you each component, with the ID of the component plasmid, component name...the component plasmids on the Addgene website. Step 4: Naming your assembled plasmid and generating...resources from Addgene Plasmids 101: Modular Cloning Addgene's CRISPR Guide Plasmids 101: Golden Gate Cloning...structure takes a lot of the design work out of the plasmid creation process. Each module of Fragmid represents...

...hot plasmids in our newsletter or on our hot plasmids webpage. Listen to the entire Hot Plasmids podcast...podcast! Hot plasmids covered in this segment Hot Plasmid 1: The Tol2 Gateway-Compatible Toolbox for quickly...In the second episode of our Hot Plasmids podcast series, you'll learn about new red fluorescent proteins...Segement on the Tol 2 Gateway-Compatible Toolbox  Hot Plasmid 2: AAV Vectors for tageting GABA-ergic interneurons...Listen to Our Segment on these new AAV Vectors Hot Plasmid 3: mCyRFP1 - a new, monomeric, and cyan-excitable... protein. Listen to Our Segment on mCyRFP1 Hot Plasmid 4: mScarlet - a new, bright, and monmeric red fluorescent...Receive Our Newsletter Check Out All of Our Hot Plasmids Articles Browse the Repository  ...

...changes we’ve made: New CRISPR plasmid pages We’ve given our CRISPR Plasmids and Resources a fresh new design...design, sorting the plasmids we have available by category. Each plasmid page now has a helpful graphic...introduce you to the plasmid class covered on that page. To help you find popular plasmids, we’ve also added... new plasmid pages that cover some of the hottest CRISPR tools. Base Edit: Base editing plasmids allow...revolution, Addgene has distributed over 100,000 CRISPR plasmids. But that’s not our only job - we strive to also...up with new developments (and, of course, find plasmids that will be useful to you.) With these goals...mind, we set out to update our web resources and plasmid pages to make them more useful and easy to navigate...

...lentiviral plasmids Addgene's most popular retroviral plasmids Addgene's collection of AAV plasmids Addgene...split across several plasmids (often three), for safety purposes. The primary plasmid with the delivered...often a gene) is called the transfer plasmid while the other plasmids with packaging or replication components...of adenovirus plasmids Resources on the Addgene blog Parts of the AAV packaging plasmid Overview of Lentiviral...For safety and production reasons, multiple plasmids with many components are used to package lab grown...can be intimidating at first to look at so many plasmid maps and protocols! In reality, there are many ...components are called helper plasmids. Below, we’ll breakdown the vector components first by virus and then...

... after ordering a plasmid you didn't have to grow the bacteria and prep the plasmid before you begin your...with 210 plasmids from the collection. You can find the full list here. Most of these plasmids are empty...cloning experiment? What if after receiving the plasmid from Addgene you could directly start your digest...you can focus on what you want to do with your plasmid without spending time and resources in preparative...research and discovery.” Here are the details. Plasmids offered as cloning grade DNA As we begin testing...molecular cloning. Plus, we decided to include 10 plasmids that can be directly used in the research on the... the SARS-CoV-2. Nine of the overall plasmid selection will be open to industry scientists. This collection...

...lentiviral plasmids include popular envelope and packaging plasmids as well as transfer plasmids. Download...Otherwise, check out our plasmid and DNA backgrounds. Download the blue plasmid background.     You can... the plasmids that Addgene offers by visiting our collections page. Here, you can browse plasmids by field...your favorite reagent, scientific technique, or plasmid repository at your next meeting.     If you’re...cute mascots!  Blugene has been helping us make Plasmids 101 videos and needed a break so they're hanging...coronavirus collection page, where you can find plasmid tools to accelerate your COVID-19 research.  Download...background.         If you’re predominantly a plasmid user, instead, don’t fret; we have backgrounds ...

...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling...gamma-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G, may be...Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral Barcoding...expressing new shRNA sequences pLKO (Plasmid #10878) This plasmid has been used by The RNAi Consortium... genes and transgene expression across several plasmids. The most commonly used viral vector systems are...popular packaging and gamma-retroviral envelope plasmids here. Lentiviral vectors are very versatile tools...guide. Popular packaging and lentiviral envelope plasmids are listed here. Please note that both lentiviruses...

...types of yeast plasmids are defined below: Yeast Integrating plasmids (YIp): These plasmids lack an ORI...our first few Plasmids 101 posts, we focused mainly on the elements required for plasmid maintenence within...not only the number of plasmids found in each cell, but also whether the plasmid gets integrated into the...integration. Yeast Episomal plasmids (YEp): These are most similar to bacterial plasmids and are considered “...simple gene replacement/mutation. Yeast expression plasmids used in the lab typically contain all the necessary...how the regulation of bacterial ORIs determines plasmid copy number within the bacterial cell. Similary...included within a yeast vector determine how the plasmid is replicated and maintained within the yeast cell...

...recommended for plasmids up to 25 kb, most cloners should be fine, especially using plasmids in the 10-12...Other Plasmid Cloning Techniques like SLIC and Golden Gate Cloning Learn How to Verify Your Plasmid Read...Reference Pages Choose Your Perfect Plasmid Backbone Find Plasmids for Your Research ...made genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While commercial kits... REPLACR (Recombineering of Ends of Linearized Plasmids After PCR). Current site directed mutagenesis...mutation, then use those to PCR amplify the template plasmid. The parental, nonmutated DNA is digested with ...of background colonies carrying the unmodified plasmid. For insertions, it’s possible to use the same ...

...important collection.  Creating the Plasmids To begin constructing the plasmids for expression of mAbs, the Trimmer...expression plasmids were DNA sequenced to determine the VH and VL sequence. These plasmids have been deposited...scientific community with the plasmid sequences available from each plasmid webpage. Importantly, the methods...Resources on the Addgene blog:  Plasmid-Based Recombinant Antibodies  Plasmids 101: Secondary Nanobody Toolbox...NABOR are recombinant antibodies expressed from plasmids that have been deposited with Addgene. Here, we... first antibodies distributed by Addgene.  The plasmids used to generate these antibodies were based off... to create an amplicon that  was cloned into a plasmid backbone for expression of the mAb heavy and light...

... AAV.  Where do you start? Plasmids of course! You just need three plasmids to start making the AAVs you...packaging plasmid which contains the AAV structural and packaging genes, the adenoviral helper plasmid which...packaging plasmid and take a look at each of its parts. Key elements of the AAV packaging plasmid The packaging...packaging plasmid aka the RepCap plasmid encodes two genes: the replication (rep) gene and the capsid (... AAV RepCap plasmid! Check out this blog post to learn more about the AAV transfer plasmid! References... for the virus to replicate,  and the transfer plasmid which contains the viral genome. In today’s blog...lab, rep and cap are expressed from a separate plasmid in trans, freeing up space in the viral genome ...

...construction of new plasmids. Dr. Duedu will also use polymerases produced using plasmids from Addgene to ... has been working with Seeding Labs to provide plasmid grants to researchers in developing countries. ... to announce that we’ve selected our first two plasmid grant awardees: Drs Louis Bengyella and Kwabena...doctors Bengyella and Duedu and how they plan to use plasmids from Addgene to advance their research. Louis ...cellulosic biomass. Dr. Duedu would like to use the plasmids he acquires through Addgene to produce molecular...fluorescent proteins. Dr. Duedu’s plans for these plasmids have a heavy focus on teaching. He hopes to teach...in environmental samples via PCR. We hope these plasmids will help Dr. Duedu inspire new Ghanaian scientists...

...a million plasmids would fill? Two weeks ago, Addgene edged passed our one million plasmids shared milestone...ending the day with 1,000,002 plasmids shipped. Reaching one million plasmids shared might have you wondering...how many plasmids leave Addgene headquarters every day. That would be an average of 673 plasmids each day...to packing and shipping. Every single day. 673 plasmids shipped daily may be easy to picture (see the ...celebratory lunch, it’s hard to picture one million plasmids. How much space would one million bacterial stabs...post, we looked at how long it takes to make one plasmid and how much time Addgene has saved researchers...Assuming 5 hours of hands-on time to make each plasmid, we would have saved 5 million hours through reagent...

...developed the plasmids, pPSU1 and pPSU2. When restriction digested with PstI or EcoRV, these plasmids generate...Penn State Ladder: Homemade Plasmid Prep Penn State Ladder: Commercial Plasmid Prep Kit Amount 1000 Lanes...Two plasmids that can be used to make inexpensive 100 bp or 1 kb DNA molecular weight ladders were recently...polyacrylamide gels. Get Tips on Verifying Your Plasmid Table 1: Comparing Typical Comericial Ladders...are significantly cheaper. When using a homemade plasmid prep protocol, it cost less than $10 to make 1000... 100 bp or the 1 kb ladder. Using a commercial plasmid prep kit increases the costs to $20-30 per ladder...prep protocol here and the pPSU1 and the pPSU2 plasmids are available from Addgene. Learn How to Analyze...

...with a plasmid that already has the multicistronic element and reporter cloned in. In these plasmids you ...interest. Some of the plasmids listed in the table above (and their related plasmids) are designed to have... a one-stop reference guide for plasmids. We've combined our Plasmids 101 blog posts and additional resources...desktop. The Plasmids 101 eBook is designed to educate all levels of scientists and plasmid lovers and ... an introduction to plasmids, allowing you to spend less time researching plasmid basic features and spend...May 11.) PubMed.   Read more Plasmids 101 posts or browse Addgene's Plasmid Guide for more molecular biology...techniques including co-transfection of two or more plasmids, the use of multiple or bidirectional promoters...