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We narrowed to 509 results for: plasmid dna

Showing: 181 - 210 of 509 results

  1. CRISPR 101: Any Base Transversion Editors

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    Blog Post
    ... inosine from DNA: N-methylpurine DNA glycosylase (MPG), also known as alkyladenine DNA glycosylase (AAG...N-methylpurine DNA glycosylase / alkyladenine DNA glycosylase  Removes hypoxanthine from DNA, creating an...create an abasic (empty) DNA site using either fused or endogenous Uracil DNA N-glycosylase (UNG) enzymes...(AP) site. UNG / UDG Uracil DNA N-Glycosylase Removes uracil from DNA, creating an abasic site. MPG...W. (2024). Programmable DNA pyrimidine base editing via engineered uracil-DNA glycosylase. Nature Communications...fused to nCas9. First, MPG removes the G base from DNA to create an abasic site. Like in most other base...non-edited strand to encourage the cell to repair the DNA using the edited strand as a template — in this case...

  2. The Stingy Scientist: How the Baby Gel Box Was Born

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    ... to make my own glass milk for purifying cut plasmid DNA from agar slices.  Molecular biologists used ...daily before the advent of techniques like PCR and DNA synthesis. Most molecular biologists bought the pricey... I realized I was pouring large agarose gels for DNA visualization or isolation, then using only a few...bromide, less stop buffer and miniscule amounts of plasmid sample.  How satisfying?  With practice, I got ...

  3. Design Tips for Prime Editing

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    ...the CRISPR guide Find CRISPR plasmids by function Browse our entire plasmid collection ... prevents the prime editor from re-engaging with DNA it has already edited. Created with BioRender.com...create 3-base (or longer) tracts of edited bases. DNA mismatch repair (MMR) is less efficient at identifying...cell to use the edited strand as a template during DNA repair. Test multiple nick sites, starting with sites... Nicking sgRNAs can be designed to target edited DNA, so that nicking only occurs after the edit is installed...genome editing without double-strand breaks or donor DNA. Nature, 576(7785), 149–157. https://doi.org/10.1038...Structure of Cas9 in Complex with Guide RNA and Target DNA. Cell, 156(5), 935–949. https://doi.org/10.1016/j.cell...

  4. Viral Vectors 101: The Retroviral Lifecycle

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    ...from a reverse-transcribed piece of DNA called a “provirus.” This DNA is synthesized inside of what is called...Central Dogma of Biology (gDNA → mRNA → protein): their roadmap includes RNA → DNA (Cooper & Temin, 1974)...transferring genes to mammalian cells (i.e. transferring DNA), yet retroviruses are RNA-containing viruses  (Robinson... because they integrate this reverse-transcribed DNA into the host cell’s genome and then use cell machinery...Transduction is often thought of as the integration of DNA into the host cell’s genome (shown in step 3), but...various mechanisms, only one of the two strands’ DNA counterparts from step 2 will be integrated by the...Hu & Temin, 1990). At step 3, the double stranded DNA is integrated into the host chromosome, and this ...

  5. Thanks to Addgene Depositors, We’re Now Sharing over 100,000 Reagents!

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    ...including plasmids, ready-to-use viral preps, pooled libraries, kits, and cloning grade DNA!     This...100,000 different plasmids available from the repository (currently at 98,276 plasmids as of April 8, 2021... 2021) 1.5 million plasmids shared in total (currently at 1,484,507 as of April 8, 2021) 5,000 depositing...addgene.org homepage! You can track the number of plasmids right below the search bar and more stats further...

  6. CRISPR 101: RNA Editing with Cas13

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    ...Learn about CRISPR DNA Base Editing Resources on Addgene.org Browse All CRISPR Plasmids Check out our CRISPR...editing has multiple advantages over more traditional DNA editing systems; first, RNA editing doesn't require...contain the RuvC and HNH domains responsible for DNA cleavage, so they cannot directly edit the genome...is mediated by ADAR2DD, not PspCas13b. Find the plasmids for the RESCUE RNA editors here! Testing and improving...for viral delivery.  Find the RESCUE RNA editing plasmids here! Future directions Building on previous RNA...Science (2017):pii: eaaq0180. PMID: 29070703. Find plasmids from this paper at Addgene. Abudayyeh, Omar O...550(7675) (2017):280-284. PMID: 28976959. Find plasmids from this paper at Addgene. Abudayyeh, Omar O...

  7. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

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    .... citrea scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines recombineering...QCascade DNA-targeting module with the crRNA, (3) three transposase proteins, and (4) the donor DNA. By using...supplied as a dsDNA fragment. Each round of editing takes two days, and the pTargetF and pCas plasmids can be...engineering functions in bacteria, but have no fear - plasmids from Addgene depositors are making it easier than...machinery to promote homologous recombination of linear DNA fragments. Since recombineering does not contain ... system available from Addgene consists of two plasmids: pCas9: carries Cas9 and chloramphenicol resistance...gRNA and a backbone-targeting gRNA cassette for plasmid curing through "self-destruction" after induction...

  8. Synthesized by Ginkgo Bioworks, Shared by Addgene: SARS-CoV-2 Plasmids for Many Expression Systems

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    Blog Post
    ...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...prepare Sanger reactions by mixing this isolated plasmid DNA with appropriate sequencing primers, and hand...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...

  9. MXS Chaining

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    ...combination of different genetic sequences into plasmid DNA. In high throughput cloning techniques, although...Gibson Assembly Learn About Other Plasmid Cloning Techniques Plasmids 101: Green Fluorescent Protein Resources...Fluorescent Protein Collection Find Plasmid Kits at Addgene Find Plasmid Cloning Protocols References 1....MXS chaining MXS-chaining was designed to create plasmids for fluorescence imaging or flow cytometry applications...modules (Figure 2) are contained in individual plasmids and are each flanked by the same multiple cloning...assembly method Single reaction, can assemble large DNA sequences, no restriction enzymes needed Not optimal...throughput cloning is to facilitate the construction of plasmids with many components. The creators of the MXS-...

  10. Plasmids 101: Codon usage bias

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    ...mutation” field on the plasmid page (see plasmid 87904 for example). As many plasmids available from Addgene...blog Plasmids 101: protein expression Plasmids 101: E. coli strains for protein expression Plasmids 101...codon optimized by ordering plasmids containing them from Addgene. If a plasmid at Addgene contains a gene...might think it’s easy to come up with a workable DNA sequence to encode your peptide of interest and produce...alternative tRNAs Codon optimization With low cost DNA synthesis, one of the primary ways researchers solve...researchers have created codon optimization algorithms and DNA synthesis companies such as IDT and GenScript host...

  11. CRISPR 101: Cytosine Transversion Editors

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    ...components: the DNA repair protein rXRCC1, or the DNA binding and lyase domain of DNA polymerase β (PB...without breaking the DNA backbone, making them efficient and precise tools for altering DNA sequences. These...structure. In order to bind DNA, Cas9 requires (1) a guide RNA, (2) a complementary DNA sequence, and (3) a protospacer-adjacent...used by Cas9 to locate a matching DNA sequence.   Target strand The DNA strand that base pairs with the...non-target strand of DNA for Cas9 to bind.   R-loop The structure formed by Cas9 binding to DNA and separating...(AP) site. UNG / UDG Uracil DNA N-Glycosylase Removes uracil from DNA, creating an abasic site. UGI...single-stranded DNA (ssDNA) region, using enzymes fused to Cas9 to alter the exposed ssDNA bases. Thus, ...

  12. Building and Validating Recombinant Antibodies: A Case Study

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    ...expression plasmids were DNA sequenced to determine the VH and VL sequence. These plasmids have been deposited...important collection.  Creating the Plasmids To begin constructing the plasmids for expression of mAbs, the Trimmer...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...

  13. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

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    ...amplify the template plasmid. The parental, nonmutated DNA is digested with methylated DNA-specific enzyme ...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...of background colonies carrying the unmodified plasmid. For insertions, it’s possible to use the same ...

  14. Tips for arabidopsis transformation

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    ....org Find plant plasmids Visit our plant resource page Deposit arabidopsis plasmids with Addgene ...pathogenic species that stably integrates transfer DNA (tDNA) into the genomes of the plants it attacks Growing...this method include: Cloning and transforming a plasmid into the bacterium Agrobacterium tumeficans - a...available (Weigel and Glazebrook, 2002). Find plant plasmid resources at Addgene  Choosing an expression system...options, but here are a couple of my favorites. Plasmids from the Tsuyoshi Nakagawa lab are binary gateway...prefer Golden Gate assembly, there are a series of plasmids from the Dinneny lab that additionally allow for...culture you are growing definitely contains your plasmid. Grow lots of Agrobacterium: The more agrobacterium...

  15. Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis

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    ...biology as the engineering of biology at the level of DNA using modular, standardized, disciplined, and open...research as can already be seen with things like DNA synthesis, Golden Gate cloning, and CRISPR. The yeast...Synthetic Biology Education Nonprofit BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at...Michael Koeris Resources at Addgene.org Find Plasmids from the Ellis Lab Find Synthetic Biology Kits...

  16. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

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    ...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...Agrobacterium-mediated DNA transfer, each GOI is usually expressed from a different plasmid, which may lead ...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...CRISPR complexes were observed binding telomeric DNA. What’s great about CASANOVA? It works with the S...

  17. Viral Vectors 101: Viral Vector Elements

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    ...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...This is often cDNA, shRNA, or something similar, and is contained on the transfer plasmid. If expression...will all be ssDNA within the viral particle. The ITRs base pair with each other to allow DNA synthesis ... the dsDNA virus. Viral packaging and entry Adenovirus is quite large and has more helper plasmid components...

  18. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

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    ... Fonfara I, Hauer M, Doudna JA, Charpentier E. A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive...genes by generating insertions or deletions in the DNA sequence. This same approach is effective to target...because very small deletions and insertions in the DNA sequence can completely disrupt expression of the... gRNAs can be used to target specific sizes of a DNA fragment and turn off the function by removing the...affect nearby genes (Figure 1A-C). Find CRISPR plasmids at Addgene! Knock-in: Introducing tags or stop... Once Cas9 generates double-strand breaks, a new DNA sequence can be knocked-in, or introduced, into the...ncRNA expression without permanently altering the DNA, CRISPR interference (CRISPRi) and CRISPR activation...

  19. History of CRISPR Cas - A tale of survival and evolution

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    Blog Post
    ...thermophilus to predatory phages to test if the exogenous DNA would be incorporated into the bacterial genome. ...specifically, specialized Cas proteins snip the foreign DNA into small fragments approximately 30bp in length...Once the CRISPR/Cas complex binds to the foreign DNA, a cut is made to destroy the invader. Wikipedia... in the field, and enabling access to multiple plasmids used for various platform applications. For more... out Addgene’s CRISPR Guide or find CRISPR/Cas plasmids for your research. Do you have a cool idea or ...

  20. Plasmids for Endogenous Gene Tagging in Human Cells

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    ...Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor plasmid contains 1 kb homology...methodology is the use of a donor plasmid that contains long stretches of DNA on either side of the fluorescent...1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into a plasmid backbone for delivery...found within the homology areas of the plasmid to prevent the plasmid from being cut and destroyed by CRISPR... the tag. Created with BioRender.com. Plasmids, plasmids everywhere To find out more about gene edited...expensive for many labs. We’ve created and tested plasmids that use CRISPR/Cas9 to endogenously tag a wide...wide variety of genes with GFP. These plasmids are available through Addgene, and the stem cell lines we...

  21. New CRISPR Tools: Cas7-11 and PASTE

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    ... that could cut RNA or DNA via a CRISPR mechanism. The weird black pellet's DNA yielded a new CRISPR effector...say the sample had their attention! They isolated DNA from the sample, amplified the CRISPR locus from ...discovery The first generation of CRISPR could cut DNA, the second generation could do base editing, and...PASTE, which can “drag and drop” kilobases’ worth of DNA with CRISPR-driven precision into a genome, can be...integrase would then integrate into with a large DNA insert – or Programmable Addition via Site-specific...take a genetic sequence and integrate it into any DNA sequence containing the 38-50 bp sequence recognized...Use PASTE can be delivered via a single dose of plasmids, works in non-dividing and primary cells, and ...

  22. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

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    ...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...instructions on how to send liquid DNA or bacterial streaks of your plasmids back to us. Our scientists will...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...

  23. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

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    ...of the second plasmid can be done in one day in one assembly reaction. This second plasmid saves you a ...alerts when plasmids become available.  We’ve also had some requests for variations on the plasmids we’ve developed...and Myers for speaking with us! Find the Plasmids: Plasmids for protein tagging with CRISPR Additional...goal for years as members of the ENCyclopedia of DNA Elements Project (ENCODE), an NIH-funded effort to... available at Addgene. For more details on the plasmids that are currently available along with detailed...system, what we are doing is adding in a second plasmid to target an epitope tag onto the transcription...second vector is a homology directed repair donor plasmid with homology arms and a selection marker. If things...

  24. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

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    ...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...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        A retroviral...transducing a only a single gRNA per cell. The viral DNA containing the gRNA then integrates into the genome...identified by next-generation sequencing of the genomic DNA. However, some cell types are poorly transduced by...treatment of cardiovascular disease. Find the LipoGlo plasmids at Addgene! Lawler et al, QJM: An International...promoter systems. Find these CRISPR activation plasmids at Addgene. Liu et al., Nature Communications,...

  25. Plasmids 101: Modular Cloning Applications and Kits

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    ...connectors, cDNAs, and more to find the perfect combination. The EcoFlex Kit has a wide array of DNA parts ...Modular cloning is a popular DNA assembly tool used to build single- and multi-gene constructs for a ...background MoClo is a clever way to assemble several DNA parts in a specific orientation using Type IIS restriction...between CRISPR components expressed from different plasmid backbones. They noted that backbone differences... as the order, or distance in the placement of plasmid elements can impact expression levels and consequently...system was used to assess how different lentiviral plasmid backbone elements impact viral titer and to compare...plants, and animals (Chiasson et al., 2019). Many plasmid elements such as promoters or fluorophores are ...

  26. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

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    ...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... our hot plasmid articles from April 2018. If you'd ever like to write about a recent plasmid deposit ...sign up here. No time to read? Listen to our Hot Plasmids Podcast! TRIM-Away: Targeted endogenous protein...segment CRISPR and RNAi have helped researchers alter DNA sequence and RNA expression - but what if you just...your protein knockdown experiments! The TRIM-Away plasmids can be found here. Clift et al. Cell. 2017. PubMed...Endosomal pH Sensors podcast segment The first Addgene plasmid deposit from Professor Stefan Hell, who won the...

  27. The Advances Behind The World's First Personalized CRISPR Treatment

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    ...-target DNA editing activity. In fact, it also reduced the (very low) levels of off-target DNA editing...Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature, 551(7681), 464–471. https...editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature, 533(7603), 420–424...was invented in David Liu’s lab by combining Cas9 DNA-targeting technology with a fused cytidine deaminase...editors did not create double-stranded breaks in the DNA backbone, making them significantly less prone to... using directed evolution to get them to work on DNA instead. However, they retained a small amount of...Analysis and minimization of cellular RNA editing by DNA adenine base editors. Science Advances, 5(5), eaax5717...

  28. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

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    ...letter of a plasmid name and simply denotes that the object is a ‘plasmid’. “p” is for plasmid. pXXXXX-XXXX...insert in your plasmid name. This is often a 3-6 letter representation of the gene (or DNA sequence). pBACKBONE-hGene...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...

  29. Editor's Choice, August 2016

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    ...can use PCR to make small sequence changes to plasmid DNA. This an oldie but a goodie, and in my personal...that describe methods to make single base edits in DNA. The first, contributed by guest blogger Kristian...experience, is a quick way to generate many mutated plasmid constructs. The second post was contributed by ...

  30. Hot Plasmids and Viral Preps - January 2021

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    ...subset of the new plasmids and viral preps 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...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...AAV-deliverable ABE.  Find the plasmid here Jesse Zalatan’s lab deposited plasmids for a chemically inducible...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll...mGold, a new yellow fluorescent protein New CRISPR plasmids New items from the viral service A fast blue-light...recognize, cut, and alter specific target sequences in DNA resulting in a variety of outcomes including excision...
Showing: 181 - 210 of 509 results