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We narrowed to 509 results for: plasmid dna

Showing: 101 - 120 of 509 results

  1. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...created DNA ladders (aka DNA molecular weight markers) that can be used to measure the size of DNA fragments...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...articles provide a synopsis of a plasmid or group of plasmids' functions and applications. We hope that... you to find and use the plasmids you need. You can find all the hot plasmids from 2017 below. With over...over 50,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...editing for precise genome modification. Plasmid ID Plasmid Name Cas9 Variant Speacial Features 85169...

  2. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...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... CRISPR plasmids are always being added to the repository. To find all of the CRISPR plasmids available...known or suspected to be involved in DNA damage response and DNA repair.  A new CRISPR pooled library ...only required three components: a master mix, the DNA of interest, and cell extract.    Graphical...They allow single-nucleotide modifications to edit DNA without requiring a double-stranded break. The Xiao...Xiao Wang Lab has recently deposited a set of plasmids that helps identify base edited cell populations...

  3. Plasmids 101: Screening Strategies Used in Plasmid Cloning

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    Blog Post
    ...once in the plasmid backbone and another enzyme that cuts within the insert. If the plasmid contains the...or absence of DNA using lysed bacterial colonies, without the need to prep your plasmid. In this case,... PCR product if the plasmid contains the insert and no PCR product if the plasmid does not contain the...precise order of nucleotides within a DNA molecule, in this case a plasmid. Sequencing is one of the surest...resources on the Addgene blog Get all the plasmid basics in our Plasmids 101 series Find more molecular biology...If you’re cloning a plasmid, you’ll need a way to find the needle in the haystack: the one perfect clone...clone that contains the plasmid you’re looking for out of the many cells that don’t. One way to begin ...

  4. Lab Automation at Addgene

    Type
    Blog Post
    ...scientists share plasmids across the globe, we’re working on ways to send plasmids as DNA as opposed to ...rate! Automating DNA purification We're developing ways to prep large quantities of DNA because some countries...more efficient. Here, I’ll discuss transformation, DNA purification, and filling vials with agar. I’ll also... transformation. With our ever growing rate of plasmid deposits (over 100 deposits per month!), manual...Biomek. The FXP id also able to do high throughput DNA preps and other molecular work if we need it to. ... shipments, we’ve needed to find a way to purify DNA from many bacterial samples at the same time as opposed...Qiagen BIOROBOT Universal Platform and it suits our plasmid prepping needs perfectly. Working with Qiagen, ...

  5. Plasmids 101: 5 factors to help you choose the right cloning method

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    Blog Post
    ... Read other Plasmids 101 blog posts New to plasmids? Read our article on What is a Plasmid? Download the...cloning different types of DNA, such as cDNA, PCR fragments, or genomic DNA. Also remember that commercially... the Plasmids 101 eBook Resources on Addgene.org0 Check out protocols for plasmid cloning Find vector... methods are more amenable to combining multiple DNA fragments than others. In today’s post, we will focus...technique, you first perform a round of PCR to generate DNA fragments with 15-40 bp of homology at their ends...combined with an exonuclease, a polymerase, and a DNA ligase to carry out a pooled set of ligations in ...enzymes, optimize reagent concentration, or purify DNA fragments from gels. If you’re relatively new to ...

  6. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...genome. The genome itself is a linear double-stranded DNA of ~36kb, with inverted terminal repeats (ITR) present...fiber, penton and hexon proteins. The viral genome DNA is further protected by histone-like proteins forming...regions and can accommodate up to 8.2 kb of transgene DNA. The second generation vectors are deficient in E1...down into 5 key steps: Construction of the rAdV plasmid (~1 week): The AdEasyTM system is the most popular...adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the transgene of interest...how to generate the final recombinant adenoviral plasmid construct, please see Addgene’s Adenoviral Guide...2.*Pro-Tip*: once the correct recombinant pAdV plasmid is identified, re-transform into a bacterial strain...

  7. Plasmids 101: The Wide World of Natural Plasmids

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    Blog Post
    ...toolbox. Natural plasmids, also called environmental plasmids, are all around us! A plasmid is a small piece...colicin-producing plasmids have been referred to as “Col plasmids” or “Col factors.” Virulence plasmids: microbial...Old Solution but a New Problem Plasmids 101: What is a Plasmid? Plasmids 101: Transformation, Transduction...F. (2023). Plasmid classifications. Plasmid, 126, 102684. https://doi.org/10.1016/j.plasmid.2023.102684...when we think of plasmids, both Archaea and Eukaryota can also harbor these circular DNA passengers. Most...., 2001) Beneficial traits of natural plasmids Natural plasmids often convey a variety of functions that...functions of natural plasmids. Created with BioRender.com. Resistance (R) plasmids: microbial defense...

  8. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    ...and was able to integrate DNA payloads of up to 15 kb. Find evoCAST plasmids here! The new horizons of...proteins integrate a payload sequence into genomic DNA downstream from the Cas target. Conveniently, the... (Tns) to integrate large sequences into genomic DNA downstream of gRNA-specified sites. Created with ...moved a gene required for phage propagation to a plasmid that lacked a promoter — only CAST integration ...20 mutations in TnsB, the subunit that catalyzes DNA insertion, along with both evolved and engineered...Transposon-encoded CRISPR-Cas systems direct RNA-guided DNA integration. Nature, 571(7764), 219–225. https://... Engineering Using CRISPR-Transposons Degrading DNA with Cascade-Cas3 Typing CRISPR Systems Prime Editing...

  9. A Look at Addgene's QC Process

    Type
    Blog Post
    ...blog Addgene's Tips for Plasmid Quality Control Plasmids 101: In-house NGS Plasmid Quality Control Now at...tools that rely on DNA amplification and manipulation to produce and edit plasmids. These are prone to...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...

  10. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ... frequent DNA repair mechanism in general. On average, it accounts for less than 10% of DNA repair, with...though exogenous DNA can be used as well. Below are some considerations for designing donor DNA for successful...homology-directed repair pathway, designing the best donor DNA, and avoiding common mishaps in this class of genome...knock-out. The goal of a knock-out is to disrupt DNA translation by generating a frameshift mutation. ...knock-ins. Knock-in mutations typically require an exact DNA sequence to be incorporated at a precise genomic ...which is done through the design and use of a donor DNA molecule. Your donor molecule will need to have homology...This homology will guide the cell to use the donor DNA as a template for repair, which will result in the...

  11. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    ... is complementary to the DNA target. The 20-nt guide region binds to the DNA sequence immediately downstream... Genome Engineering Guide Plant Plasmids and Resources CRISPR Plasmids and Resources ... types, it has a limited capacity to carry large DNA cargos. As a result, the search for smaller, compact...single RuvC-like domain. In simple terms, Cas9 cuts DNA using two "scissors" (one for each strand), while...PAM (protospacer adjacent motif) near the target DNA, TnpBs also need a short sequence motif called a ...reRNA) or omega RNA (ωRNA) guide the TnpB to its DNA target, much like the single guide RNA (sgRNA) used...downstream of the TAM. This RNA-DNA hybrid formation recruits TnpB to the site and activates its nuclease...

  12. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...Chromatin is a complex of DNA and histone proteins that compacts cellular DNA into chromosomes and reduces...reduces the risk of dangerous DNA damage. If the way DNA is wrapped around histones changes, then gene expression...methyl- groups to the DNA, which mostly occurs at CpG islands (regions of the DNA with a high frequency...gene expression that do not involve any changes to DNA sequence (Epigenetics literally means ‘on top of ...This is how most cells in the body contain the same DNA but have very different structure and function, and...change as well. The more frequently and tightly bound DNA is to histones, the more compacted it becomes. This...associated with reduced life-span, but no changes in the DNA sequence could be found. Angelman syndrome and Prader–Willi...

  13. Viral Production at Addgene

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    Blog Post
    ...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral...transfer plasmid Addgene uses a rigorous barcode matching system that follows the sample from the DNA tube.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...confirmation of transfer plasmids used in Addgene’s viral production. Barcoding allows our DNA and virus to be ...of the viral genome and the absence of unexpected DNA contaminants. The data is also used to determine ...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...lentiviral vector, incubated for 72 hours, and genomic DNA is extracted. Primers and probes targeting integrated...

  14. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...Mobile-CRISPRi ICE element forms a conjugative plasmid-like DNA circle and is transferred to recipient bacteria...Browse our CRISPR interference plasmids Find bacterial CRISPR plasmids Read our CRISPR guide ... into the genome and is propagated during normal DNA replication and cell division. Mobile-CRISPRi does...transposon system. This system has two parts: a plasmid expressing the Tn7 transposition genes (tnsA, tnsB... tnsB, tnsC, tnsD, and tnsE), and a plasmid that contains the Mobile CRISPRi cassette flanked by left ...copy of the RP4 transfer machinery mobilize these plasmids for transfer to recipient cells during a tri-parental...due to site selection by TnsD, a sequence-specific DNA binding protein. Selection removes donor bacteria...

  15. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...primers flanking my edits in genomic DNA and doing PCR on genomic DNA. The surveyor assay itself was relatively...BRDN0000561167) (Plasmid #80034) and BRAF gRNAs (BRDN0001145345 and BRDN0001146266) (Plasmid #76479 and #76480...that Surveyor nuclease cleaves double stranded DNA (dsDNA) at basepair mismatches. To use this tool to ... denature and renature the dsDNA. Upon renaturation, you will generate DNA duplexes with one strand from...viruses, but you can find a list of validated gRNA plasmids on our website. There are a few reasons to target...gRNA-transduced Cas9-A549 cells are used as a control. DNA was analyzed by PAGE. The basis of the Surveyor...these mismatches are indicated by the presence of DNA cleavage products. The appearance of the 428 bp band...

  16. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...reflection of the large size of the plasmid or silencing of the plasmid by DNA methylation. Another downside...into cells as a donor plasmid containing the transposon, along with a helper plasmid that expresses the ...reprogramming factors. Non-replicating Plasmids: Generating iPSCs with plasmid-based expression requires serial... generating iPSCs by plasmid transfection where 2 of 11 clones tested had plasmid integration. Another...the dose of plasmid the cells receive over the whole reprogramming period and the plasmid will get diluted...larger plasmids have lower rates of transfection. Minicircles: Minicircles are like mini-plasmids. They...periods of time than traditional plasmids due to lower activation of DNA-silencing mechanisms. While minicircles...

  17. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ... consists of a Ti plasmid, into which the GOI is inserted, and a Ti "helper" plasmid, which contains Vir...necessary for T-DNA processing and insertion into the plant genome. B) The Ti plasmid and helper are transformed... to plant cells. The region of the Ti plasmid between the T-DNA borders (yellow) are recognized by the...for creating plant expression plasmids, including plant CRISPR plasmids from the Yiping Qi lab and the...plant-specific modifications to commonly used CRISPR plasmids are necessary to use the CRISPR system in plant...et al., 2013). Addgene carries Cas9-containing plasmids for knockout, activation and repression of target...expression of both Cas9 and the gRNA off of the same plasmid. Delivering CRISPR components to plant cells Once...

  18. What's New in CRISPR - September 2019

    Type
    Blog Post
    ...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...scaffold and a domain needed for DNA unwinding. They’ve deposited plasmids for active and inactive CasX for...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...Jennifer Doudna’s lab discovered a new Cas protein, CasX, from metagenomic analysis of microbial DNA from ...In this system, Cas9 generates a double-stranded DNA break at the genomic loci of interest and on either...vector sequence. Homology arms that flank the donor plasmid cargo are complementary to either side of the double-stranded...expression. Read the paper in Nature Find the CasX plasmids Drug inducible transcriptional activation and...

  19. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...complementary DNA (cDNA), and then use PCR and molecular cloning to insert the cDNA into the desired plasmid backbone...experiments in planarians with their recent plasmid deposit. These plasmids enable two main types of experiments... cloned into plasmids to investigate for each of these planarian species. These plasmids all have a PR-T4...dsRNA. Students can produce DNA templates via PCR off of the same plasmid and use these to create riboprobes...off of linearized DNA templates which they can easily generate off of a circular plasmid via PCR. Typically...Resources at Addgene.org Find plasmids from the Sánchez lab Find plasmids for other developmental models...of specific gene expression. The 49 deposited plasmids enable students to study gene expression and functions...

  20. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...to the genomic DNA of interest  Figure 1. When ABEs are in action they bind DNA so that the TadA...Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464–471 . https://doi....TadA domain can edit the short single-stranded DNA stretch that is accessible. Image from Gaudelli et al...in 40 new ABE8 variants. When comparing on-target DNA editing efficiency in a human cell line for these...ABE8.x-m/d constructs demonstrated a 2.1x on-target DNA-editing efficiency in a human cell line while reducing...pyogenes Cas9 requires the target site in the genomic DNA to contain a specific protospacer adjacent motif ...followed by 2 guanines). This limits the use of ABEs as DNA sites that do not contain a suitable PAM cannot be...
Showing: 101 - 120 of 509 results