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We narrowed to 893 results for: plasmid

Showing: 201 - 220 of 893 results

  1. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...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...

  2. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...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.        AAV delivery... Figure 1: The base editor is split into two plasmids and co-transfected into cells for reconstitution...overcome this overlap, Yiyang Gong’s lab created two plasmids containing a red-shifted Ace-mScarlet, a fusion...channelrhodopsins. Find the Ace-mScarlet voltage indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed...fragments. This system has been used to construct plasmids for use in model organisms such as E. coli and...

  3. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ...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...write about a recent plasmid deposit please sign up here.     Listen to the Hot Plasmids episode! Expanding.... Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid as well as several...further probe cellular function. The CRY2clust Plasmid can be found here. Park et al. Nat Commun. 2017..., the Barrick lab has deposited the pDCAF3-IS5 plasmid containing a synthetic operon that enables the ...the ΔguaB pDCAF3 strain. Find the decaffeination plasmid here. Quandt et al. ACS Synth Biol. 2013. PubMed...

  4. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...single plasmid transformation, though, a pooled library has hundreds to millions of distinct plasmids, all...anyone who has worked with individual plasmids knows, some plasmids amplify well and others don’t. The size...pooled library, which contains a collection of plasmids in a single tube, or a pre-made viral prep library... prepare the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral...library and appropriate packaging and envelope plasmids into HEK cells allows production of lentivirus...amplify the library. This is similar to amplifying a plasmid: transform E. coli with the pooled library DNA,...own protocol that amplifies without disrupting plasmid representation. A good place to start is with a...

  5. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...bacteria with the plasmid can grow. As the bacteria divide, they replicate and retain the plasmid. This allows... Inside Addgene Posts! Learn all about Plasmids in our Plasmids 101 series! ...Every plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the...about Developing Lab Management Software    The plasmid's journey begins First, each package is opened and... to hundreds of samples, with an average of 10 plasmids per deposit. Each week an average of 16 deposits...allow us to store multiple types of tube for each plasmid we have at Addgene. Each sample at Addgene has ...strains that have been treated to easily take in plasmids (transformation). Transformed bacteria are then...

  6. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ... that made it easy to share plasmids, both for people depositing plasmids and for people requesting them... Collections Lentiviral Plasmids Tetracyline Inducible Plasmids Empty Backbones Bacterial Expression...Expression Systems AAV Plasmids Blogs Origin of Replication What is a Plasmid? How to Design Your... Verify Your Plasmid Using a Restriction Digest Analysis Videos What is a Plasmid? How to Design...video, however, is “What is a Plasmid?”, An animated explainer of a plasmid and its parts that helps the...them. Prior to this, many plasmids were available, but they often weren’t easily accessible, due to a ...barriers, allowing people to access a wide catalog of plasmids from all over the world. This idea, that available...

  7. Plasmids 101 eBook 4th Edition

    Type
    Blog Post
    ...popular eBook, Plasmids 101, is now available!   Plasmids 101 covers everything from plasmids parts and ...word for it!  “When people want to learn about plasmids, they don’t go to Wikipedia; they go to Addgene...

  8. Hot Plasmids and Viral Preps - May 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...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...Hutchinson Cancer Research Center developed a pooled plasmid library encompassing all possible amino acid mutations...anti-SARS-CoV-2 antibodies. In the library, each plasmid encodes the mutated spike protein and a barcode...head over to our CRISPR Plasmids and Resources page.  A new system uses light to deactivate photocleavable... accumulation of indels to measure time. These plasmids with different promoters driving expression of...

  9. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...Guide Pages Find CRISPR Plasmids for Your Experiments Deposit Your New Plasmid Tools with Us  ...Integration into Target Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about... knock-ins, some labs have used NHEJ for whole plasmid integration; however, since NHEJ is error-prone...cloning. An overview of PITCh. The PITCh plasmid contains a GFP-Puro cassette flanked by 5' and ...sites. This PITCh plasmic is cotransfected with a plasmid carrying Cas9, the PITCh-gRNA, and the locus-specifc.... The PITCh-gRNA creates two DSBs in the PITCh plasmid to release the cassette, and an insertion-locus-specific...insertion. PITCh can also be adapted for whole-plasmid integration if you’d like to integrate a larger...

  10. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...Addgene’s plasmids are used with a wide variety of restriction enzyme-based cloning methods. Each method...Entry DNA overhangs may be present in the original plasmid (Option 1) or added using PCR-based amplification...fragments to create TAL effector nucleases. These plasmids were designed to utilize the BsaI and BsmBI Type... a gRNA target sequence into a Cas9-containing plasmid such as pX330. This cloning strategy not only makes...makes it easy to create a single gRNA-expressing plasmid, but it can also be adapted to express multiple...create CRISPR/Cas9 constructs, assemble standard plasmids parts in different combinations, or other new ...PubMed. More Resources at Addgene:  Check out more Plasmids 101 Posts! Check out Addgene’s restriction cloning...

  11. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...light-responsive system is spread across three plasmids, with the response regulator encoded in the genome...et al. condensed the system to two streamlined plasmids. The utility of these TCSs was limited by a few...These bulky systems were spread over a number of plasmids, with one RR (ompR) encoded chromosomally. Leakiness...constructs have been greatly streamlined; only two plasmids are required per system. Applications of these...many strains of E. coli, as well as with other plasmid-based systems, which will enable the assembly of...sb500273n. Epub 2014 Oct 11. PubMed. Find the plasmids from this publication at Addgene. Synthetic biology...2005 Nov 24;438(7067):441-2. PubMed. Find the plasmids from this publication at Addgene. A synthetic...

  12. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    ...effector nuclease (TALEN) kit consisting of 834 plasmids. The researchers showed they could use their TALEN...particular DNA sequence? Musunuru: The kit includes 832 plasmids containing prefabricated pieces that allow you...the genome in rapid fashion. The two remaining plasmids in the kit have the rest of the protein in place...fluorescent proteins used to sort the cells after the plasmids have been introduced by transfection or electroporation...protein that binds those 15 base pairs is take four plasmids out of the library in the appropriate combination... ligation with the four inserts and the master plasmid in one step. The kit also has other unique aspects...you are designing a TALEN on paper to a finished plasmid. With our protocol, it is feasible to produce genetically...

  13. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ....12137. PubMed. Find the Plasmids at Addgene Kowarz Lab plasmids Izsvak Lab plasmids  ...the gene of interest (blue) in a plasmid backbone. A separate plasmid contains the transposase gene (red...). The original plasmid is empty following the removal of the transposon; the plasmid is then degraded...for insertion in the correct orientation. These plasmids use a novel synthetic RPBSA promoter, which is...

  14. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

    Type
    Blog Post
    ...individual lentiGuide gRNA plasmids targeting the human kinome. These plasmids contain the same 3-4 gRNAs...true biological insight. Pooled and individual plasmid formats A first for a library at Addgene, you can...first subset of the pooled library. The individual plasmids will be useful when screening for phenotypes, ... screening in a pooled setting. The individual plasmids will also be useful for imaging or gene expression...assays necessitating the screening of individual plasmids separately. Additionally, this set can be used... Page Find Validated gRNAs Deposit Your CRISPR Plasmids ...

  15. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...cytosine demethylation in mammalian cells. This plasmid is used with pcDNA3.1-MS2-Tet1-CD to decrease methylation...Tet1CD, is available from Rudolf Jaenisch’s lab in plasmid form or as ready-to-use lentivirus. Figure...Staphylococcus aureus Cas9 (Sa-Cas) and deposited plasmid IF311: pMAGIC (R4-R3) NLS-Sa dCas9-NLS-LSD1. LSD1...experience with these constructs in the comments! Find Plasmids for Epigenetic Modification   Leah Schwiesow...510–517 . https://doi.org/10.1038/nbt.3199 Find plasmids from this publication at Addgene. Klann TS, Black...561–568 . https://doi.org/10.1038/nbt.3853 Find plasmids from this publication at Addgene.  Lei Y, Zhang...8: . https://doi.org/10.1038/ncomms16026 Find plasmids from this publication at Addgene. Liu XS, Wu ...

  16. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    ...gRNA-containing plasmids (McDade et al, 2016). Many of the gRNAs contained within these plasmids have been ...scientists’ submissions, either their deposited gRNA plasmids or from submission of a spreadsheet containing... validated gRNAs that are not generated from a plasmid. If you, for example, prefer to deliver your CRISPR...columns are target gene, species, sequence, Addgene plasmid ID (if we have one associated with the gRNA), application...please consider submitting their sequences (and plasmids!) so that this shared resource can continue to... Tools Read Our CRISPR Guide Pages Find CRISPR Plasmids  ...

  17. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...expression plasmid encoding the ORF to be expressed Finally, the researcher needs a plasmid encoding their...#68306 pCRT7 NT TOPO E17TAG GFP plasmid - Addgene #68295 SupD plasmid – Addgene # 68307 For further technical...Find Synthetic Biology Plasmids Find Bacterial Expression Systems Find Plasmids for Protein Purification...strain for protein production, and an expression plasmid encoding the open reading frame (ORF) of interest...translation of pSer-incorporated proteins on a single plasmid. This includes 4 copies of a tRNA that specifically... during translation. Many ‘flavors’ of Sep-OTS plasmids are available through Addgene, but we have worked...assay using the PhosTag system (see below). This plasmid backbone can also be cloned with an ORF of interest...

  18. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...that the plasmid could be maintained in the absence of antibiotics, resulting in the plasmid pBAD-bARGSer-AxeTxe... 1248 ng of CMV-gvpA-IRES-BFP plasmid and 954 ng CMV-gvpNJKFGWV plasmid into 70%-confluent well of HEK... Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram of pBAD-bARGSer-AxeTxe. (b) Methods to ...overexpressed compared to accessory proteins. Excess gvpA plasmid added into the transfection mixture in HEK293T ...mammalian species to produce gas vesicles. New mARG plasmids Our new mARGs come as a set of two PiggyBac integratable...genes gvpNJKFGWV marked with Emerald GFP. These plasmids can be used directly in transient transfection... into the genome by cotransfecting them with a plasmid expressing PiggyBac transposase. In rtTA-cell lines...

  19. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    ...Best Way to Elute and Store Your Plasmid DNA? Selecting Your Plasmid Purification Kit Resources on Addgene.org...You have just purified your plasmid and are ready to move on to your downstream application — but wait...wait! Do you know how much plasmid DNA you have or how pure your sample really is?  To assess the concentration...concentration and purity of nucleic acids, such as plasmids, the go-to method is to use spectrophotometry....Protein contaminants Whether you are purifying plasmid DNA from bacteria or cleaning up a PCR product,... 230 nm is guanidine thiocyanate (used in many plasmid purification kits), which can dramatically reduce...assay or running an agarose gel. In the case of plasmids, the use of RNase treatment to degrade bacterial...

  20. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ... involving two-plasmid or single-plasmid systems (Vo et al., 2020). In the two-plasmid system, an effector...group’s initial report of VchINT, three separate plasmids and multiple promoters were used to express the...effector plasmid (pEffector) expresses all seven required genes and the CRISPR RNA under the same promoter...promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also be combined...combined in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and ...and the ability to easily cure cells of the plasmid after integration. Figure 2: (A) Streamlined ...insertions with Type I-F INTEGRATE using a single plasmid containing a multi-spacer CRISPR array. Workflow...
Showing: 201 - 220 of 893 results