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We narrowed to 509 results for: plasmid dna

Showing: 271 - 300 of 509 results
  1. Rett Syndrome: A History of Research and Therapeutic Outlooks

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    Blog Post
    ... that a small DNA binding motif in MECP2 interacts with the minor groove of AT-rich DNA sequences to regulate...their MECP2-related plasmid tools with Addgene and share additional resources. Plasmid tools to facilitate...to partner with them to launch a Rett Syndrome Plasmid Collection and Resource Center for Rett Syndrome... Rett Syndrome in novel ways.  Addgene’s MECP2 Plasmid Collection and Resource Center will continue to...
  2. An Introduction to Adenovirus

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    ...., 2019). AdVs are non-enveloped double-stranded DNA viruses, with a linear genome between 26–45 kb in...initiating the whole process – such as the viral DNA polymerase – it makes sense that vectorization strategies...strategies include creating an AdV transfer plasmid with the E1 and E3 (Early 1 and Early 3) genes deleted.... These genes’ functions are rescued by helper plasmids used during production and/or by genes present...made available by this deletion from the genomic plasmid can be used to house a recombinant sequence (transgene...late genes are usually present in the transfer plasmid for the majority of adenoviral vector systems used...
  3. CRISPR 101: Multiplex Expression of gRNAs

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    ...multiple gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the...rectangles) are cloned into various plasmids using oligonucleotides. These plasmids contain Type IIS restriction...different purposes.   Gersbach Lab multiplexing plasmids: This plasmid set allows you to express 2-4 gRNAs, with...assembled to produce a plasmid encoding the gRNAs and Cas9. As with the Gersbach lab plasmids, multiple Cas9 ...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids allow you to assemble 2-4 gRNAs through...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids can be used to express up to 8 gRNAs after...Yang Lab Single Transcript Multiplexing Plasmids: These plasmids are similar to the Csy4 polycistronic ...
  4. Keeping Up With CRISPR/Cas9

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    Blog Post
    ...s incredible.” Looking for CRISPR Plasmids? CRISPR-Cas9 Plasmids available at Addgene CRISPR-Cas9 References...Forbes respectively: "A Powerful New Way to Edit DNA" and "This Protein Could Change Biotech Forever."...Last month, the Lawrence Berkeley Lab's Jennifer Doudna and Eva Nogales presented the first detailed picture...
  5. Open Enzyme Collection

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    ...Synthetic Biology Plasmids BIOFAB Plasmid Set Drew Endy Lab Plasmids Jennifer Molloy Lab Plasmids Blog: Golden...assembly. Plasmids Explore the collection and order individual plasmids via the links below: DNA Ligases... ID Plasmid Gene/insert 165503 pOpen-HSlig4 DNA Ligase IV 165514 pOpen-9N7lig 9 Degrees North DNA Ligase...pOpen-Pfulig Pfu DNA Ligase 165535 pOpen-THEAQlig Taq DNA Ligase 165554 pOpen-T4gene30 T4-DNA Ligase 165564...pOpen-ECOligA E. coli DNA Ligase 165569 pOpen-Tthlig Tth DNA Ligase 165574 pOpen-HSlig3 DNA Ligase III Return... to top DNA Polymerases Addgene ID Plasmid Gene/insert 165500 pOpen-9N7polA 9 degrees N-7 DNA polymerase...Tli DNA polymerase (exo-) 165524 pOpen-T4gene43 T4 DNA Polymerase 165532 pOpen-ECOpolA E. coli DNA Polymerase...
  6. Sequencing Options for CRISPR Genotyping

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    ...the edited strand and wild-type strand of the host DNA. When these strands hybridize, the nuclease can detect...account other elements, including those relating to DNA structure (e.g. epigenetic modification, bulges) ... et al. "High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity...Frock, Richard L., et al. "Genome-wide detection of DNA double-stranded breaks induced by engineered nucleases... Crosetto, Nicola, et al. "Nucleotide-resolution DNA double-strand break mapping by next-generation sequencing...Addgene.org View Our CRISPR Guide Pages Find CRISPR Plasmids for Your Experiments Get Ready-to-Use CRISPR Lentivirus...
  7. Improved Plasmid Maps Powered by SnapGene

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    ...SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid maps far easier...easier for you to share plasmid information with colleagues, or explain plasmid features to a lab member...sequences, & downloadable .gb & .dna files Once you’ve identified a plasmid that you want to learn more about... on the main plasmid page under the "Resource Information" section. Analyze your plasmid sequence For ...over from our users: "Please, please improve your plasmid maps!" After thoughtful design, vetting, and tweaking...tweaking, we’re excited to announce that our plasmid and sequence displays are now powered by GSL Biotech's...software and extensive feature library, our updated plasmid and sequence displays are now much easier to interpret...
  8. Analyzing CRISPR Editing Results with ICE from Synthego

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    ... of CRISPR components into target cells, genomic DNA from both edited and unedited (control) populations...from single or multiplex CRISPR-Cas9 double-strand DNA breaks using SpCas9. More features will be added ...the user-defined 17-23 nucleotide sequence of the DNA-targeting region of the guide RNA, excluding the ...Addgene.org Check out our CRISPR guide Find CRISPR plasmids for your research Find other CRISPR software resources...
  9. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

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    Blog Post
    ...independently searched for this unique sequence of DNA and both identified lymphocyte antigen 6 complex ...receptor for PHP.B by looking for a unique sequence of DNA that was present in mice whose CNS was transduced...chromosome. Of the genes located in this stretch of DNA, Ly6a seemed the most likely candidate due to its...permissive and non-permissive mouse strains. PHP.B plasmids and PHP.eB viral preps are available from Addgene...
  10. Plasmids 101: Inducible Promoters

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    ...Resources on the Addgene Blog: Plasmids 101: The Promoter Region Plasmids 101: Cre-lox Light-Switchable...protein, the repressor protein is removed from the DNA. With the repressor protein absent, transcription...Promoter Subtype Promoter Example Activator Addgene Plasmid/Kit Alcohol-regulated AlcA promoter AlcR pGGA008...use light to regulate transcription. Red flame plasmid pDawn contains the blue-light sensing protein YFI...Systems for E. coli Browse other blog posts from our Plasmids 101 series Resources on Addgene.org: Inducible...Inducible Systems at Addgene Cre-lox Guide and Plasmids     ...
  11. 15 Years of Addgene: The Top 15 Plasmids

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    ...and an envelope expressing plasmid, such as our second most requested plasmid of all time, pMD2.G. Find...wide range of applications. This plasmid was the top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB... top requested plasmid from 2016. You may notice the “V2.0” at the end of this plasmid which indicates...prior version of this plasmid. Indeed the first version is the 15th most popular plasmid of all time, but ...envelope expressing plasmid pMD2.G. Find pMDLg/pRPE. scramble shRNA - This plasmid from David Sabatini....1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...Cas9, this plasmid expresses Cas9n (a D10A nickase mutant) to nick, rather than cleave DNA. Find pSpCas9n...
  12. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

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    ...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...
  13. Overcoming the AAV Size Limitation for CRISPR Delivery

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

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    ...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 ...
  15. Handling Plasmids from Addgene - Purifying Plasmid DNA

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    ...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...
  16. Fluorescent Proteins 101: Fluorescent Biosensors

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    ...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...
  17. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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

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    ...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 ...
  19. SciComm: Taking Science to Elementary and Middle Schools

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    ...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 ...
  20. Plasmid Tools for Microbiome Studies

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    ...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...
  21. Plasmids 101: Broad Host Range Plasmids

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    ...Addgene blog Plasmids 101: Origin of Replication Plasmids 101: What is a PlasmidPlasmids 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...
  22. Plasmid Cloning by PCR (with Protocols)

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    ... 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...
  23. CRISPR Guide

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    ...-stranded DNA (dsDNA) plasmid. Small single-stranded DNA (ssDNA) oligos often work well for small edits...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...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...
  24. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

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

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    ...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...
  26. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

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

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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...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...
  28. CRISPR Plasmids - Bacteria

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

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

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    ...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, ...
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