... successfully takes up plasmid DNA. Among those that do, some now have plasmids that contain your gene...including ccdB on its destination plasmids. In the destination plasmid, the ccdB gene is flanked by recombination...selections in plasmid cloning. Additional resources on the Addgene blog Read our Plasmids 101 blog posts...purify your gene of interest, get it into your plasmid backbone, and zap the mixture of DNA into cells...gene of interest, but others will uptake plasmid backbones that re-ligated back on themselves. Therefore...strategy needs to identify cells containing the plasmid construct you’re seeking. Fortunately, there are... example, if the DNA you’re inserting into the plasmid backbone contains a kanamycin resistance cassette...

... Bioblasts? Plasmagenes? In the 1940s and 50s, scientists were working to understand genetic cytoplasmic...how did plasmids get their name? In 1952, Joshua Lederberg set out to clarify the classification of these...these cytoplasmic inheritance factors. He proposed the catch-all term “plasmid” derived as a hybrid of ...tetracycline resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102, with EcoRI...engineering, cloning, and plasmid tags, addtional content added to the "Plasmids that Glow" and "Viral Expression...Learn more about the history of plasmids and cloning: CSHL Meeting: Plasmids: History & Biology The Joshua...of "cytoplasm" and "id" (Latin for 'it'), as “a generic term for any extrachromsomal hereditary determinant...

...letter of a plasmid name and simply denotes that the object is a ‘plasmid’. “p” is for plasmid. pXXXXX-XXXX...information about plasmid features and tips you can use in the lab, check out more posts in our Plasmids 101 series...There are no universal rules for naming plasmids but here are some good guidelines to follow in order...people can quickly and easily identify what your plasmid contains and other important information. Tip:...Backbone name Include the empty backbone name in your plasmid name. This simple piece of information can often...important details. Once you know the backbone a plasmid is based on, you can usually derive: a) the bacterial...  Include information about the insert in your plasmid name. This is often a 3-6 letter representation...

...2017 and last updated Apr 6, 2021 by Will Arnold. Introducing NGS in 2017 for all incoming plasmids was a...every isolated plasmid sample and normalize each to fall within a 10 fold range. Plasmids that don't meet...A broader view of the whole plasmid To confirm the sequence of the plasmid, we examine three things: ...function of the plasmid.   Confirming the insert We usually confirm the insert through BLAST or by direct...expected and do not affect plasmid function, we will update information on the plasmid’s page. Sometimes we’re...’re missing a piece of the plasmid There are regions of some plasmids that are particularly difficult ...some plasmids will not result in one complete, circular assembly. Most of the time, these plasmids are ...

...the plasmid based on the predicted sizes and organization of different features within the plasmid. Restriction...if you don't have the full plasmid sequence. Once you have purified plasmid DNA, this method can be done...example plasmid on the right has a total size of 7.3kb, including a 1.2 kb insert. The plasmid was digested...possible plasmid conformations, with relaxed and nicked marked with asterisks (*). When the plasmid is digested... Congratulations, you have a plasmid expressing your gene of interest (YGOI) and are ready to dive into...functional experiments! Whether you’ve cloned the plasmid yourself or obtained it from a colleague down the...with the correct construct, and verify that the plasmid you received matches the expected sequence. Here...

... metabolic enzymes including biotin carboxylase and decarboxylases. These enzymes catalyze the transfer...biotinylating proteins with BirA. You can find plasmids that contain both AviTag and BirA at Addgene! ...

... on All Things Plasmid with Our Plasmids 101 Series Get Tips on Verifying Your Plasmid Resources on Addgene.org...Check out Our Plasmid Reference Pages Deposit Your Completed Plasmids Find the Perfect Plasmid for Your Research...recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid Bonus...recipient plasmid. Remember to insert your DNA in the correct orientation in the recipient plasmid by viewing... of DNA so that it can be easily cloned into a plasmid of interest. You can use similar processes to add... Interest (YGOI) for ligation into a recipient plasmid. Designing primers for PCR based cloning   ...this example). Assuming you are amplifying from plasmid DNA (rather than from genomic DNA or a cDNA library...

... (blue) light switch. Zhang’s team devised the plasmids now available in Addgene’s repository for use ...biological processes and disease states. While LITE plasmids have clear application to anyone with an interest...the new optogenetic tools in C. elegans and in Plasmodium. For those who do want to apply LITEs for the...

...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!) ... Cas13. CRISPR-Csm degrades both nuclear and cytoplasmic mRNA with 90-99% knockdown efficiency across ... genes and RNA-seq of a nuclear (MALAT1) and cytoplasmic (CKB1) RNA target. Adapted from Figure 3 Colognori...Colognori et al 2022.   Find CRISPR Csm plasmids here! Colognori D, Trinidad M, Doudna JA. Precise transcript...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...

...Addgene blog:  Plasmids 101: What is a plasmid? Plasmids 101: Origin of ReplicationPlasmids 101: PCR...process known as plasmid multimerization. Plasmid Multimerization Early studies on plasmid topology recorded...ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form of the plasmid. Have... monomer of Plasmid 12259: pMD2.G (depicted on the left) is ~5.8 kb in length. The plasmid can exist on... have been utilizing full plasmid sequencing services to verify the plasmids, including long-read sequencing... the sequencing results for their plasmids, they found the plasmids were much longer than expected, and...tandem. Such plasmids are known as multimers, and they are the result of multiple plasmid copies combining...