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We narrowed to 508 results for: dna plasmids

Showing: 281 - 300 of 508 results

  1. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...analyzing the plasmid and viral DNA sequences and expression profile at multiple steps from plasmid amplification...to flip back into the inverse orientation (See Plasmids 101: FLEx Vectors). While this strategy greatly...recombination that are typically employed during viral plasmid amplification (Fischer, KB. 2019). This means that...of this finding, we now include guidance on the plasmid page of DIO/FLEx AAV vectors where this occurs ... mechanism of spontaneous reversion during AAV plasmid amplification. On the left homologous pairs of ...Recombinant Adeno-Associated Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678. https://....2020.618020 Additional resources on the Addgene blog Plasmids 101: FLEx Vectors Important Considerations When...

  2. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ...bacterial immune system cleaves bacteriophage and plasmid DNA. Nature 468:67–71 . https://doi.org/10.1038/nature09523...capabilities of full-length SpCas9. This set of plasmids includes AAV-Cas9C-VPR for targeted gene activation...Structure of Cas9 in Complex with Guide RNA and Target DNA. Cell 156:935–949. https://doi.org/10.1016/j.cell...Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids ..., Staahl BT, Taylor KL, Desmarais J, Nogales E, Doudna JA (2019) CasX enzymes comprise a distinct family...

  3. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    ...this case, plasmid copy number is the input and gene expression is the output. The input, plasmid copy number...to decouple gene expression and plasmid number. In a suite of plasmids modified from pSC101 with copy ...of all types of plasmids, the stabilized promoters were also tested in different plasmid backbones that...stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, ...RepA is a plasmid replication initiator protein, they could alter copy number of the plasmid by altering...genome or a plasmid. Thus, a stabilized promoter can allow a system to be moved from a plasmid to the genome...special collections page Download the Plasmid 101 eBook See all plasmids deposited by the Voigt lab ...

  4. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...Recovering Plasmid DNA from Bacterial Culture Recovering Plasmid DNA from Bacterial Culture You may also...concentrated plasmid DNA. This page will discuss the general procedure for purifying plasmid DNA from bacterial...high concentration of clean plasmid DNA. To improve the purity of plasmid DNA purified without a kit it ...isopropanol to precipitate the plasmid DNA. Either spin to pellet the DNA or apply the solution to a column...How to purify plasmid DNA from a bacterial culture, including protocols, tips, and FAQ Protocols... new window) Promega sell kits for isolating plasmid DNA in quantities as low as a few micrograms to as...describe the general procedure for purifying plasmid DNA from bacterial cultures. If you will be using...

  5. Fluorescent Proteins 101: Fluorescent Biosensors

    Type
    Blog Post
    ...Resources on Addgene.org Find Biosensor Plasmids Find Optogenetics Plasmids Visit the Fluorescent Protein Guide...associated publication for the specifics of your chosen plasmid. Types of biosensors When choosing a biosensor...biosensors to follow processes such as protein or DNA binding, enzyme or transcriptional activation, conformational...

  6. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...γ-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G (Vesicular...in trans by specific plasmids. This method, therefore, reduces the number of plasmids that are required ...encoded by the transfer plasmid, but instead are provided in trans by other plasmids or the packaging cell...do many γ-retroviral plasmids have similar names? The nomenclature of the plasmid backbone is meant to...components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA of interest..., which facilitate integration of the transfer plasmid sequences into the host genome. Typically, it is... genome upon viral transduction. Most transfer plasmids available from Addgene were derived from MoMLV...

  7. Safe Port in a Storm...How Addgene is Weathering the Pandemic

    Type
    Blog Post
    ...urgent requests for access to coronavirus-related plasmids and to receive important coronavirus-related deposits...shipping staff agreed to come in and start getting plasmid deposits into the collection and to start shipping...support Addgene now, when their distribution of plasmids for COVID-19 research is accelerating essential...’ve recently piloted Addgene’s new cloning grade DNA service, and viral vector distribution is recovering...-related slowdown. More scientists are sharing plasmids and other reagents every day, and this crisis ...

  8. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    ...the same page in terms of the vocabulary--DNA, protein, plasmid, gene, etc. Though I talked about CRISPR...kid’s face when you’ve shown them how to isolate DNA from a strawberry, or demonstrated that they’re living...out our comprehensive CRISPR guide Find CRISPR plasmids and resources ...

  9. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...conjugative mobilization of broad host-range IncQ plasmids." Plasmid 62.2 (2009): 57-70. PubMed PMID: 19465049...page Deposit microbiology plasmids with Addgene Find synthetic biology plasmids ...family-RP4 conjugation system. This system can deliver plasmids to both Gram-positive and Gram-negative cells.... Wang lab further developed a modular suite of plasmids (pGT) with various replicative origins that can...genera across four phyla that have taken up the plasmid through conjugation in vivo studies. However, they... tools for microbiome studies.  Find the MAGIC plasmids here Paper-based synthetic biology approach to...difficile. As mRNA expression cannot be identified with DNA-based qPCR methods, the ability to detect mRNA transcripts...

  10. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...Addgene blog Plasmids 101: Origin of Replication Plasmids 101: What is a Plasmid?  Plasmids 101: Transformation...host range in plasmids? Not all plasmids were created equally. Only a handful of plasmids have been identified...range plasmid with only one ori and a broad host range plasmid with multiple oris that vary in DnaA-box ...broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001  Meyer...define plasmid host range, identify a few of the features that confer broad host range in plasmids in nature...recombinant vectors.  A plasmid’s host range is determined by the different hosts that a plasmid can transfer to...influence the plasmid’s ability to establish and maintain itself within host cells.  The plasmid has multiple...

  11. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ... you are amplifying from plasmid DNA (rather than from genomic DNA or a cDNA library), roughly 18-21bp...may also like... Restriction Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary PCR based...and NotI to ligate our cDNA into the recipient plasmid. Remember to insert your DNA in the correct orientation... Protocols Plasmid Cloning by PCR Plasmid Cloning by PCR You may... piece of DNA and at the same time adding restriction sites to the ends of that piece of DNA so that it...recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid. Bonus.... Digest Your DNA Set up restriction digests for your PCR product and recipient plasmid. Because you lose...

  12. CRISPR Guide

    Type
    Collection
    ...purify specific regions of DNA, image DNA in live cells, and precisely edit DNA and RNA. The ease of generating...Browse Plasmids: RNA Targeting , RNA Editing Degrading DNA with Cas3 Rather than initiating DNA cleavage...-stranded DNA plasmid. Small ssDNA oligos often work well for small edits like a single-point mutation...homology with the target DNA. Once the Cas9-gRNA complex binds a putative DNA target, the seed sequence...will begin to anneal to the target DNA. If the seed and target DNA sequences match, the gRNA will continue...strands of the target DNA. This results in a double-strand break (DSB) within the target DNA, located ∼3–4 nucleotides...of target DNA. Cas9n increases target specificity. Since two nickases targeting opposite DNA strands are...

  13. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    ...This construct will serve as the repair template plasmid to create fluorescent and 3xFLAG-tag fusion proteins...proteins. The repair template and Cas9-sgRNA plasmid is then injected into the germline of young adult ....115.178335. [Epub ahead of print] PubMed. Find plasmids from this publication at Addgene. Other strategies...10.1038/nmeth.2641. Epub 2013 Sep 1. PubMed. Find plasmids from this publication at Addgene. Scalable and... and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans...genetics.114.170423. Epub 2014 Sep 23. PubMed. Find plasmids from this publication at Addgene. Additional...

  14. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...You may also like... Restriction Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary The...promoters, selectable markers, or any other DNA element between plasmids. Let's assume that you are beginning...recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid Will...sites to your recipient plasmid : You can modify the MCS of your recipient plasmid using Annealed-oligo ...Procedure Digest your DNA Set up restriction digests for your donor and recipient plasmids. Because you lose.... We recommend 1.5-2μg of donor plasmid and 1μg of recipient plasmid. It is also critical that as much...re-circularization of the recipient plasmid. You should treat your digested recipient plasmid with a phosphatase prior...

  15. 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...easily assemble multiple DNA fragments. This system has been used to construct plasmids for use in model organisms...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...

  16. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ... Protocols Restriction Digest of Plasmid DNA Restriction Digest of Plasmid DNA You may...... Recovering Plasmid DNA from Bacterial Culture Purifying DNA from an Agarose Gel DNA Ligation Introduction...digest of plasmid DNA Protocols...Electrophoresis chamber Pipetman Reagents Liquid DNA aliquot of your plasmid of interest (see below for recommend ...to digest your plasmid. Pro-Tip To determine which restriction enzymes will cut your DNA sequence (and ...typically involves ∼500 ng of DNA, while molecular cloning often requires 1 µg of DNA. The total reaction volume...volume of DNA to be cut. A typical restriction digestion reaction could look like this: 1 µg DNA 1 µL of...

  17. CRISPR Plasmids - Bacteria

    Type
    Collection
    ...efficiency than NHEJ. ID Plasmid Gene/Insert Promoter PI Publication Browse CRISPR Plasmids By Function Genome...cytosine base editor that can alter DNA bases without inducing a DNA break. Cytosine base editors convert...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert Promoter... purify a region of genomic DNA and its associated proteins, RNA, and DNA. The enCHIP system uses an anti-FLAG...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In...Ampicillin none, need Cas9 plasmid Qi Do you have suggestions for other plasmids that should be added to ...

  18. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    ...receive transfer plasmid DNA that you can use along with lentiviral packaging plasmids to create your own...rough approximations of the total number of CRISPR plasmids for that particular organism available at Addgene...helped distribute many of these technologies in plasmid form and are proud to have facilitated their fast...fast adoption. However, in many cases the plasmids themselves are only the starting point for the production... use; after receiving a pooled gRNA library in plasmid format, a researcher must amplify the library, ...

  19. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...the Video! DNA Ligation Assemble DNA using DNA ligase Bacterial Transformation Introduce DNA into a bacterial...phenol-chloroform extract, and precipitate DNA DNA Quantification Measure DNA concentration with a spectrophotometer...and analyze plasmid DNA with restriction enzymes Agarose Gel Electrophoresis Separate DNA by size on an...Add a short stretch of DNA to a plasmid Gibson Assembly Combine overlapping DNA fragments in a single ...Store bacterial strains or plasmids for long term use Watch the Video! DNA Purification Miniprep, phenol-chloroform...Amplify CRISPR pooled-plasmid libraries Diagnostic Restriction Digest Verify a plasmid using restriction ...FAQ Watch the Video! Purifying DNA from an Agarose gel Isolate specific DNA bands cut from a gel Watch the...

  20. Meet the New Addgene Board Members!

    Type
    Blog Post
    ...over 100 requests for the plasmids and the lab eventually deposited these plasmids at Addgene. While deposits...she says. In 11th grade she heard a talk about Z DNA that inspired her to major in molecular biology in...The mission to share critical reagents such as plasmids and viral stocks for researchers around the world...
Showing: 281 - 300 of 508 results