...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...

...all of us here would add materials to that list (plasmids, yay!). Scientists are blogging, talking to the...

...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...

...Heidelberg University. “What is different is the plasmids you use to bring this into your target organism...

...don't exactly "eat" plasmids as depicted in this comic, when a bacterium uptakes a plasmid, the genes on the... the plasmid can give them different phenotypes. For example, this plasmid encodes a green fluorescent... Read all of our Plasmids 101 blog posts Check out all of our blog posts on plasmid cloning Resources...comes from adjacent lysed bacteria and can include plasmid DNA or fragmented DNA released into the environment...membrane so the bacteria can easily uptake your plasmid of interest. Scientists can also use electroporation...exist as transient extrachromosomal DNA, like a plasmid, or it can integrate into the host bacterium’s ...cloning vector that contains both bacteriophage and plasmid properties. The phagemids are packaged into replication-incompetent...

...with some plasmids. You just need three plasmids to start making AAV:  the packaging plasmid which contains... transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids have two ...of an AAV transfer plasmid. Parts shown in green are key elements of an AAV plasmid. Parts shown in grey... elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal ...transfer plasmids have a promoter, but this element isn’t required when the transfer plasmid harbors non-coding...Transfer Plasmid video!   Now you’ve got a basic understanding of some common AAV plasmid elements!...structural and packaging genes, the adenoviral helper plasmid which contains the proteins needed for the virus...

...Addgene “Find Plasmids” badge in the image below. If an abstract on PubMed has the “Find Plasmids” badge, you...you’ll find new plasmids. Simply click on the badge to be taken directly to the plasmids on Addgene’s site...you should see the “Find Plasmids” box on papers that have deposited plasmids. Now that you know how ...Order Plasmids from Addgene Check Out Our FAQ Pages Browse Plasmids ...thought - Is there an easy way to find published plasmids? Do you spend a lot of time in PubMed searching...you can use PubMed to see which articles contain plasmids deposited at Addgene from the abstract page. Check...article to ensure you understand the features of the plasmids, how they were created, and how to use them in...

...hold the plasmid together, even if there are gaps or flaps remaining. This incomplete plasmid looks like...cloning uses short sticky ends to insert genes into plasmid backbones, base pairing between these ~4 base overhangs... overhangs isn’t strong enough to hold a plasmid together through transformation, so the cloned vector...longer single-stranded overhangs can hold a nicked plasmid together long enough to get it into cells and let... of LIC is often limited to specially designed plasmids. Sequence- and ligation-independent cloning: A...fixing up the “damage” and producing a complete plasmid. Besides being compatible with any vector, the ...into bacteria, which will efficiently repair the plasmid. Created with BioRender.com.   SLIC also opened...

...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...

... 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...

...curated collections of plasmids as kits with greatly reduced costs per plasmid. We generally create kits...just the tool you need. The plasmids in this portion of the BIOFAB Plasmid Set contain terminators of ...populations. This kit consists of a 48 plasmid adapter library, with each plasmid carrying a unique barcoded Illumina-M13F...researcher at a nonprofit institution, you can order plasmids covering a wide range of fields from Addgene. ...kits to allow you to easily use a set of plasmids together on a tight budget. To help popularize our ...order from the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit... production (a variety of pExpression vectors) plasmids. The type IIS restriction sites used in these ...

... 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 ...

...underrepresented plasmids (plasmids that were lost or “dropped out”) and overrepresented plasmids (plasmids that ...In addition to single plasmids, Addgene also distributes pooled plasmid libraries containing hundreds...library contains many plasmids, care must be taken to ensure that all of these plasmids are amplified and ...diversity. If the various plasmids in the library are differently sized, smaller plasmids may be amplified preferentially...hundreds, thousands, or even a million plasmids. These libraries are some of Addgene’s most exciting and versatile...harvested, much like one would prep an individual plasmid. However, lots of library DNA is needed for screening...100 to 1,000 bacterial colonies per individual plasmid in the library. Bacterial growth must also be optimized...

...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...

...you encounter mycoplasma contamination. Causes of mycoplasma contamination The mycoplasmas enter the cell...removal of mycoplasma but, can get rid of most of mycoplasma contamination and are the last alternative... growth of mycoplasmas. The serum is nutrient-rich and also the best source for mycoplasma proliferation...suppress the growth of mycoplasma but cannot eradicate it completely. When mycoplasmas are seen in the culture...highly expensive). Detecting mycoplasma contamination Detection of mycoplasma by naked eyes or optical microscope...most of the mycoplasma species, apart from a few of them. Another method used for mycoplasma detection ...infecting mycoplasmas are used. A broad range of primers encompasses almost all the infecting mycoplasma. The...

...Not all plasmid preps are the same. Before purifying a plasmid from a bacterial culture, it is important...of purity. Based on these premises we can classify a plasmid preparation in 3 different ways: Transformation... into a plasmid backbone. Transfection grade DNA Figure 3: Need to introduce plasmids into mammalian... an external plasmid in mammalian cells and it will likely require micrograms of plasmid DNA for each ...larger scale, to allow the extraction of enough plasmid to last for multiple experiments. Another fundamental...utilize your plasmid DNA, so that you can decide what will be the optimal type of plasmid preparation ...Addgene blog Browse blog posts on plasmid cloning Read our Plasmids 101 blog posts Learn about the differences...

...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 ...Does it make a difference? Short answer: yes. This last step will influence the stability of your sample...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 ...

...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 ...

...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate...30, 2020 and last updated Jul 12, 2021. What started out as a small collection of plasmids for coronavirus...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...

...We've developed a new plasmid toolkit, the Broad Institute Target Accelerator Plasmid Collection, that we...scale laboratory-based functional experiments that classify which variants are likely gain-of-function, loss-of-function...correlations between variants in a gene of poorly classified function and a well established cellular signaling...phenotypes. Broad Institute Target Accelerator Plasmid Collections To overcome this first challenge, we...developed the Broad Institute Target Accelerator Plasmid Collections, now widely available to the scientific...in our recent publications, we used the cancer plasmid collections listed above to demonstrate how gene... hope that by releasing these new and exciting plasmid collections (together with the data generated from...