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We narrowed to 237 results for: Mammalian expression plasmids

Showing: 1 - 20 of 237 results

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

    Type
    Protocol
    ...bacterial expression vector. Using subcloning, you can easily move YGOI into a mammalian expression vector...YGOI for short). You might need to express YGOI in cultured mammalian cells. The problem is that the only...recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid Will...correct orientation in the recipient plasmid. (You don't want to express the antisense version of your gene...sites to your recipient plasmid : You can modify the MCS of your recipient plasmid using Annealed-oligo ...material. 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...

  2. General Transfection

    Type
    Protocol
    ...Use this protocol to transfect mammalian cells with your plasmid of interest....transfecting mammalian cells using linear polyethylenimine. Transfections allow for transient expression of a ...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ..., the plasmid should also be propagated in an endonuclease negative E. coli strain such as NEB stable....is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1–2 ...the cells 24–48 h before checking for protein expression. Sample Data Legend: Lenti-X 293T cells were ...here by the amount of green fluorescent protein expression (green in the right panels) with a limited effect...

  3. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl serum-...to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio that gives...drives expression of puromycin. Puro R Puromycin resistance gene for selection of pLKO.1 plasmid in mammalian...shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G Envelope plasmid for producing viral... also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG from Robert Weinberg...into the pLKO.1 vector, producing a final plasmid that expresses the shRNA of interest. C.1 Recommended ... transient knockdown of protein expression, you may transfect plasmid DNA directly into the target cells...

  4. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    ...discernable effect on mammalian cells. In this blog post we will discuss how mammalian plasmids differ from their...their genomes. Unlike plasmids used for expression in yeast and bacteria, plasmids used for stable transfections...creating stable cell lines. Looking for Mammalian Expression Plasmids? Check Out Addgene's Empty Backbone ...Your Perfect Plasmid Backbone Read More: Plasmids 101: Antibiotic Resistance Genes Plasmids 101: Origin...Although plasmids do not naturally exist in mammals, scientists can still reap the benefits of plasmid-based...synthetic vectors and cultured mammalian cells. Of course, these mammalian vectors must be compatible with...replication (ORI) will not allow for plasmid replication in mammalian cells, for example, and a toxin that...

  5. Plasmids 101: Protein Expression

    Type
    Blog Post
    ... system. Mammalian expression systems Mammalian cells are an ideal system for the expression of mammalian...designed for mammalian expression utilize viral promoters (SV40, CMV, and RSV) for robust expression post-transfection...culture conditions similar to mammalian expression systems. Bacterial expression systems When one wants to...on Addgene.org Bacterial expression systems Search for plasmids by expression system ...biology in our Plasmids 101 eBook! Researchers use various techniques to control protein expression for experimental...types of expression systems used for protein production and purification. These include mammalian, insect... and cell free expression systems. Overall the general strategy for protein expression consists of transfecting...

  6. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...Description Expression Additional considerations CMV General expression mRNA Strong mammalian expression promoter...some cell types. EF1a General expression mRNA Strong mammalian expression from human elongation factor ...consistent expression regardless of cell type or physiology. SV40 General expression mRNA Mammalian expression...Analogous to mammalian EF1a promoter. GDS General expression mRNA  Strong yeast expression promoter from...lower expression than U6. May have better expression in neuronal cells. U6 small RNA expression shRNA...promoter, which is necessary for getting the plasmid express our gene of interest. This element is responsible...enhancer. PGK1 (human or mouse) General expression mRNA Mammalian promoter from phosphoglycerate kinase ...

  7. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...TLDR: This is a versatile mammalian expression vector that you can use to express both SpCas9 and your gRNA...gRNA in a variety of mammalian cell types. Purpose  SpCas9 This plasmid expresses Streptococcus pyogenes...Similar to last year, the top requested plasmid from 2016 is an expression vector for CRISPR nuclease, SpCas9...for SpCas9 expression post transfection by western blot. Overall, this is a versatile mammalian expression...you can use to express both SpCas9 and your gRNA of interest in a variety of mammalian cell lines. Alternative...Read Some Common Ways to Deliver CRISPR Plasmids to Mammalian Cells Resources on the Addgene Website ...for sure, as is obvious from our most requested plasmid, SpCas9 is still going strong as the basis for ...

  8. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... the most desirable level of expression. Over time, transgene expression in a polyclonal population may...Protocol to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector...protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral...lentiviral vector. Unlike the short-term protein expression observed using transient transfection approaches... lentiviral vectors enables long-term protein expression studies. Moreover, repeating experiments in a...transfection. Some lentiviral vectors deliver mammalian antibiotic resistance (e.g., puromycin, blasticidin...check the cells regularly to confirm transgene expression. As polyclonal populations of resistant cells...

  9. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...packaging plasmids as well as transfer plasmids. This concept ensures safe handling and expression of these...cloning and expressing your gene of interest with GFP expression.  Cloning and expressing RNAi pMKO.1 ...researchers to express siRNAs of their choice in mammalian cells.  Learn more about Mammalian shRNA Tools...separating viral genes and transgene expression across several plasmids. The most commonly used viral vector...gamma-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G, may be...gamma-Retroviral Transfer Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector.... CRISPR technology pdCas9-humanized (Plasmid #44246) Expression of a catalytically inactive (dead), human...

  10. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...pAquaN1 Cyan Mammalian Expresses mammalian optimized Aquamarine mCerulean N1 Cyan Mammalian Express a gene...pCEP4CyPet-MAMM Cyan Mammalian Expresses mammalian optimized CyPet pCyPet-His Cyan Bacterial Expresses CyPet with...Yellow Mammalian Expresses mammalian optimized YPet pYPet-His Yellow Bacterial Expresses YPet with C-...interest: Plasmid Color Expression Description pPROEX Aqua Cyan Bacterial Expresses Aquamarine with...Yellow Mammalian Express a gene of interest fused to the C-terminus of SYFP2 Clover Green Mammalian Expresses...of monomeric Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus...with C-terminal His tag SCFP3A Cyan Mammalian Express a gene of interest fused to the C-terminus of SCFP3A...

  11. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...or a fusion protein. Mammalian vectors One of our most popular mammalian expression vectors, c-Flag pcDNA3...as close as possible to the basal expression level, induce expression at an opportune time or go all out...pcDNA series. It has the CMV promoter for mammalian expression, a multiple cloning site to insert YGOI,.... You can also upgrade your basic mammalian expression system and go for a lentiviral or a retroviral ... following the expression pattern of a defined promoter region, ectopically expressing YGOI in a pattern...you find what you need.   More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular...for overexpressing YGOI in an isolated, controlled environment. In most cases bacterial expression vectors...

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

    Type
    Blog Post
    ...greyscale.   ARG expression in mammalian cells The first-generation mammalian ARGs (mARGs) enabled... needed to increase expression. We were able to express Anabaena GVs in mammalian cells without the shell...constitutively-expressed mARG plasmids driven by CMV promoters. Labs that are new to the mARG expression are advised... that, when expressed, enable cells to be visualized under ultrasound imaging. Expression of ARGs results...on bacterial ARGs and Tom focused on mammalian ARGs. ARG expression in bacteria The first-generation bacterial...in bacteria, expressed poorly at body temperature (37oC) and were too burdensome to express in situ. As...that the plasmid could be maintained in the absence of antibiotics, resulting in the plasmid pBAD-bARGSer-AxeTxe...

  13. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...System Application Mammalian expression vector Promoter driving Cas9 expression can be constitutive ...we’ll provide an overview of some CRISPR mammalian expression systems, the typical applications for each...donor plasmid or single-stranded DNA oligonucleotide, so you will need to make sure your expression system... components into your target cells.  Mammalian CRISPR expression systems Each model system has its own...included with each expression system as well as suitable applications. Expression System Components ... lines. Transient or stable expression of Cas9 and/or gRNA in a mammalian cell line that can be transfected...particles. Stable, tunable expression of Cas9 and/or gRNA in a wide variety of mammalian cell lines. Useful for...

  14. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...natural mammalian plasmids, mammalian vectors are even less similar to bacterial ones. The mammalian analogue... Resistance Gene Plasmids 101: Yeast Vectors Plasmids 101: Mammalian Vectors Plasmids 101: The Promoter...other plasmids? If we all use the same molecular code, can I transfect any old bacterial plasmid into ...lines that express viral machinery. Alternatively, some slow-growing cells, particularly mammalian cells like...blog Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection Plasmids 101:...bacteria first. In fact, most yeast vectors and many mammalian vectors are designed to shuttle between species...used across species: replication, selection, and expression. Replication It’s usually not enough to get your...

  15. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ...changes we’ve made: New CRISPR plasmid pages We’ve given our CRISPR Plasmids and Resources a fresh new design...design, sorting the plasmids we have available by category. Each plasmid page now has a helpful graphic...introduce you to the plasmid class covered on that page. To help you find popular plasmids, we’ve also added... new plasmid pages that cover some of the hottest CRISPR tools. Base Edit: Base editing plasmids allow...breaks. Our new page shows you base editing plasmids for mammalian systems, bacteria, plants, and yeast. Popular...contains Cas13 constructs for mammalian and bacterial systems. Plasmids with wild-type Cas13a (formerly...Genotyping Day 5: Delivery systems: CRISPR 101: Mammalian Expression Systems and Delivery Methods and A Match...

  16. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ... be used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even plants...conditional gene expression, multiplexed CRISPR, and chromosomal integration of larger expression cassettes ...chromosomal integration and gene expression, the system includes a toolkit to express recombinant proteins within...interactions in gene expression, and to generate multi-gene knockouts. Mammalian and fly applications ...developed for non-mammalian systems, but it is not confined to that arena! The Mammalian Toolkit was engineered...specifically for the types of expression and integration needed in mammalian cell types, and comes with ...great option for mammalian cloning, and is geared towards cloning multi-TU expression constructs (Martella...

  17. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...vectors and an envelope expressing plasmid, such as our second most requested plasmid of all time, pMD2.G....PX459) V2.0 - This plasmid from the Zhang lab is a mammalian vector that expresses both SpCas9 and your... the envelope expressing plasmid pMD2.G. Find pMDLg/pRPE. scramble shRNA - This plasmid from David Sabatini...(PX461) expresses both a Cas9 protein and a gRNA. But instead of Cas9, this plasmid expresses Cas9n (a...generation lentiviral packaging plasmid from the Didier Trono lab that expresses the HIV gag, pol, rev, and...Didier Trono lab is pMD2.G, a VSV-G envelope expressing plasmid. Envelope proteins determine which types ...wide range of applications. This plasmid was the top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB...

  18. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...to be expressed. These permit exquisite control of gene expression and tracing of gene expression across... basal expression. Co-expression of repressor QS along with QF leads to intermediate expression from QUAS...transgene permits directed expression of that gene in cells that also express GAL4. Usage of this system...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...basal state, and co-expression of the inducer QF can increase QUAS driven expression by ~3,300-fold in ...Gossen M, Bujard H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters...cumate gene-switch: a system for regulated expression in mammalian cells. BMC Biotechnol 6, 43 (2006). https...

  19. Hot Plasmids - August 2020

    Type
    Blog Post
    ...Addgene #140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using...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...marking microtubules of mammalian cells, the author found that too high expression of these markers can ...They generated plasmids tagged with EB3 to mark the growing tips of microtubuli and plasmids tagged with ...microtubules. Find these fluorescent mammalian cell structure markers plasmids at Addgene! Chertkova, AO, et ...easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed in the cytosol...

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

    Type
    Blog Post
    ... be easily expressed in non-mammalian cells. Yeast expression plasmids The yeast expression constructs...cleavage, codon usage, and expression system. Mammalian expression plasmids Finally a set of wild type...S. cerevisiae expression vectors are available at Addgene. A set of mammalian expression vectors containing....  Bacterial expression plasmids For those constructs to be used in bacterial expression systems, Ginkgo...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...Ginkgo synthesized SARS-CoV-2 expression plasmids in a high copy backbone with a T7 promoter driving multiple...
Showing: 1 - 20 of 237 results