Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 921 results for: Tro;

Showing: 391 - 420 of 921 results

  1. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...qPCR assays, samples, standard curves, and normal controls must be run in triplicate.  The nice thing about... of DNA in your sample relative to your normal control - you can set up your calculations so that your...so forth. This also serves as a light technical control - an RQ value of 1.5 means you have either made... does not require a standard curve or a normal control, it can take a few hours longer than qPCR to run...AT-rich DNA and by variation in amplicon length. Am J Trop Med Hyg. 2008 Dec;79(6):887-92. PMID: 19052298. ...

  2. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    ...preimplantation embryos that are produced during in vitro fertilization treatments and donated afterwards ... produced Ethical concern High. Embryos are destroyed during ESC isolation. Low Chances of post-transplantation... the reprogramming factors were delivered via retroviral infection [8]. The work was taken forward the...generation of human iPSCs from dermal fibroblasts via retroviral and lentiviral infections respectively [9, 10...cell types. Like ESCs, iPSCs can differentiate in vitro into derivatives of all three germ layers, form ...

  3. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...same result – uncontrolled movement of the vehicle or, in the case of cancer, uncontrolled cell proliferation... cellular level there is a universal theme - uncontrolled cellular proliferation. We have appreciated ...appreciated for some time the causes of this uncontrolled proliferation, in particular the roles of proto-oncogenes...opposite task, keeping cells from dividing out of control. One may compare the roles of proto-oncogenes and...

  4. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...of all distributed plasmids as part of quality control, but does not perform functional tests. The newest...produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA requires incubation at 55...can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA...does however rely on cloning into a vector or in vitro synthesis, which could be less desirable if one ...et al. "DNA-guided genome editing using the Natronobacterium gregoryi Argonaute." Nature biotechnology ...

  5. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...drug-resistant population compared to a non-treated control group. In this scenario, gRNAs that are “enriched...future therapeutic targets for diseases causing uncontrolled cell growth, such as cancer. Figure 2...As with any experiment, the use of appropriate controls, multiple replicates and several cell types can...of multiple gRNAs targeting the same gene can be strong evidence that a particular gene is actually important...Titering Your Lentivirus Plasmids 101: NGS Quality Control for Pooled Libraries Find More CRISPR Resources...

  6. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...techniques involve the assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation...specifically cuts methylated DNA, so it will selectively destroy the template, but not newly synthesized sequences...protein-coding sequence, or a mutation can be introduced in a gene, while adding a GFP-tag (Figure 3)....unamplifiable’ Check your amplification by gel electrophoresis. In our experience, good amplification after...interested in neuronal signal transduction, and uses electron cryo-microscopy to understand receptor function...

  7. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...diseases such as hemophilia, muscular dystrophy, spinal muscular atrophy, neurodegenerative diseases and metabolic...life-threatening (cystic fibrosis, hemophilia, muscular dystrophy)? AAV gene therapy started in the mid-90's with...larger transgenes. Unfortunately it has limited tropism in the retina, primarily infecting cells of the...epithelium (RPE). AAVs, on the other hand, have broad tropism in the eye. AAV-2, the first identified serotype...

  8. MXS Chaining

    Type
    Blog Post
    ... create a single polycistronic 15 kb insert. The final construct was then introduced to HeLa cells (Figure...resulting HeLa cells show robust labeling, with strong expression and detection of each fluorophore at...meaning that (in the previous example) one can control whether the third module is added to the 5’ terminus...degree of identity, so not suited to generate polycistronic containing multiple identical polyadenylation...

  9. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...now many variations, including many that use auxotrophic rescue genes as reporters and some that repress...example of the former. In this system, histidine-auxotrophic host strains are used to select for binding-dependent...accounted for using appropriate replicates and controls (see Tips below), and by validating leads with...reporter gene activation. Including a prey-only control also provides a baseline level of reporter activation...that are known to bind serves as a good positive control to ensure that your setup works. The last thing...

  10. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    ...generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP system ...requested plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene...Addgene: Your lab recently developed a retroviral enChIP system using CRISPR technology. Can you describe...Associated with an IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. Fujita...

  11. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...research. Many other mutations have since been introduced to GFP and new iterations of fluorophores are... Transcription Reporter: Placing GFP under the control of a promoter of interest can be used to effectively...Scientists can use light to detect, measure, and control molecular signals, cells, and groups of cells in...Plasmids of this type may have the GFP under the control of an additional promoter from that of the gene...fate studies. It can also be used, when put under control of promoters of interest, to visualize the developmental...

  12. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ...editing has quickly become a popular system for in vitro and germline genome editing, but in vivo gene editing...injury markers were not increased compared to a control AAV-GFP. Not only did the AAV-SaCas9-sgRNA constructs... mediates protein trans-splicing, similar to an intron in pre-mRNA splicing. In 2016, Fine et al developed... L, Heidenreich M, Banerjee A, Habib N, Li Y, Trombetta J, Sur M, Zhang F (2014) In vivo interrogation...Toolbox Read about CRISPR enzyme Cpf1 Read our Introduction to AAV Resources on Addgene.org Check out ...

  13. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...axis determined by the researcher, based on their controls. You can usually tell where this point is by looking..., M., & Al-Qahtani, M. (2018). Evaluation of in vitro chondrocytic differentiation: A stem cell research...Minguela Puras, A., Pons, J. A., & Muro, M. (2018). In vitro intracellular IFNγ, IL-17 and IL-10 producing T ..., Li, J., Greiner, D., & Anderson, D. (2015). Neutrophil Responses to Sterile Implant Materials. PloS ...Plasmid-based Recombinant Monoclonal Antibodies Introduction to Antibodies  ...

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

    Type
    Blog Post
    ... situ. As a result, they had to be expressed in vitro before the bacteria were injected, limiting their...toxicity or burden relative to a fluorescent protein control. We also added the toxin-antitoxin stability cassette...visible under both phase contrast microscopy and electron microscopy (Fig. 1b). Gas vesicle expression ...batch-sorted to isolate a polyclonal cell line with strong gas vesicle expression and robustness without chromatin-opening...Biomolecular Tools for Acoustic and Magnetic Imaging and Control of Cellular Function. Biochemistry 56, 5202–5209...

  15. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ... you’d first introduce your sgRNA sequence into the pTargetF construct and then introduce both plasmids... genome edits but the double strand breaks it introduces makes it difficult to perform two or more integrations...cutting of the plasmid. At this point, subsequent introductions of pTargetF containing another sgRNA targeting...expression which can be unstable and lack precise control over copy number. MUCICAT system contains 4 plasmids...

  16. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...factors. The episomal vectors are introduced into cells by electroporation. hiPSC colonies are obtained by...to perform cell therapy and to model diseases in vitro. Find stem cell plasmids at Addgene Many sources...Fibroblasts need to be cultured and expanded in vitro for several passages to produce an adequate number... by 10 days after electroporation, most of which are integration-free, karyotypically normal and are able...

  17. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...DNA dot blot...Transduction assay...SDS-PAGE...Electron microscopy… Any guesses? That's right... ways...genome-containing viral capsids is typically quantified by electron microscopy of a viral vector solution. Since this... this technique is intensive and requires an electron microscope, it is not routinely performed on all...genome-containing (Figure 1).    Figure 1: Electron micrograph of an Addgene AAV viral vector preparation...Considerations When Using AAVs Addgene's AAV Quality Control Methods Check Out Our Viral Vector Featured Topic...

  18. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...development and evolution of histone genes in Strongylocentrotus purpuratus, the spiny purple sea urchin. ...that they form including genetic mechanisms that control who a female wants to mate with and if she decides...Lab (@EvoEmbryo) September 24, 2018   Xenopus tropicalis (Gary Gorbsky) This is not a new experimental... pan-arthropod genetic manipulation. The ReMOT control system (Receptor-Mediated Ovary Transduction of...

  19. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...organisms to control their movements and behavior. These functions and others are controlled by collections...tomography, super-resolution STORM imaging, and electron microscopy. Sample preparation for these techniques... et al. "High-performance probes for light and electron microscopy." Nature methods 12.6 (2015): 568. ...the Addgene Blog Rabies and neuronal tracing Retrograde AAV: Making the journey from axon to nucleus ...

  20. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...Christina Mork, Jul 27, 2020; updated by Susanna Stroik January 24, 2023. DNA damage drives genome instability...direct ligation of sticky or blunt ends and often introduces small insertions and deletions along the way....the original DNA sequence at the DSB, or it can introduce specific mutations into the damaged DNA if these...repairing your target DNA until a mutation is introduced disrupting one or both of these sequences. To...Easi-CRISPR (Quadros et al., 2017). Easi-CRISPR in vitro transcribes RNA encoding your repair template, then...

  21. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...for with in vitro display methods such as phage display (Bradbury et al., 2011). In vitro selection avoids...expressed as scFvs and screened with in vitro assays to select for strong binders. Then the sequences for the...2011) Beyond natural antibodies: the power of in vitro display technologies. Nat Biotechnol 29:245–254 ...

  22. Five Popular Model Organisms

    Type
    Blog Post
    ...easily control the level and spatiotemporal expression of a gene of interest. This type of control over ... (Zuber et al., 2009). By injecting mice with retroviral vectors that harbored different mutations found... modular vectors that can be used to optimally control gene expression in specific cells of the fly (Pfeiffer...using HOTcre, a method that uses heat induction to control the temporal expression of different transgenes...

  23. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...beta-barrel structure. Although the beta barrel has no strong affinity for other proteins, it may still block...There are several ways to measure stickiness. In vitro, ultracentrifugation or gel filtration is the most...Avoiding protein over-expression artifacts The introduction of DNA that encodes a fusion protein adds protein...knock-out of the corresponding protein, that is strong evidence that the fusion behaves well. But this...fusion proteins: random insertion of GFP with an in vitro transposition reaction." BMC neuroscience 3.1 (2002...

  24. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...which refers to mEGFP. This variant was made by introducing two mutations (F64L, S65T) in the original jellyfish...fluorescent proteins can be accurately determined in vitro by measuring the fluorescence intensity of purified...Peptide-tags for site-specific protein labelling in vitro and in vivo." Molecular BioSystems 12.6 (2016): ... "Blue fluorescent amino acid for biological spectroscopy and microscopy." Proceedings of the National...Additional Resources on the Addgene Blog Read our Introduction to GFP Read our Fluorescent Proteins Blog Posts...

  25. Finding Your Perfect Job After University

    Type
    Blog Post
    ...least 3 months experience working in a developing tropical country. This is why I set up a placement working...very hard to speak with individuals who had very strong beliefs in traditional medicine, especially when...very new, and everything was more structured and controlled. I worked from Standard Operating Procedures ... reliant on programming skills which are not my strong point. Working in industry: virology On completing...because it is an English speaking country with a strong economy and plenty of jobs in science at world ...

  26. Top Requested Plasmid of 2017 - pMD2.G

    Type
    Blog Post
    ...lentivirus envelope plasmid pMD2.G from Didier Trono’s lab! Lentivirus envelope vector pMD2.G This plasmid...expresses the VSV-G envelope protein under the control of the powerful mammalian CMV promoter. Envelope...protocols and information, be sure to check out the Trono Lab website. If you have plasmids you’d like to ... Guide to All Things Lentivirus 5 Tips for Troubleshooting Viral Transductions Getting the Most from Your...

  27. New Viral Vectors - Summer 2024

    Type
    Blog Post
    ... AAV9 Controls Jensen New serotype pAAV-CAG-2xNLS-tdTomato-WPRE-hGHpolyA AAV9, PHP.eB Controls Feng ...from the Gradinaru lab. This is useful for both in vitro and in vivo work with many cell types, as CAG is...serotypes pAAV-hSyn-GRAB_OTmut AAV9 Biosensors (Control) Li New viral service pAAV-hSyn-GRAB_ACh3.0 AAV9...

  28. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...FITS In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However...However, in vitro screens cannot faithfully model the complex spatial, cellular, metabolic, and cytokine...modified cells; and (5) is compatible with the introduction of unique molecular identifiers, which can improve...includes pLX_305_Ovalbumin, a plasmid that enables introduction of the model antigen, Ovalbumin, into cell lines...

  29. AAV Packaged on Request is Here!

    Type
    Blog Post
    ...offer serotypes AAV1, AAV2, AVV5, AAV8, or AAV Retrograde (AAVrg), and volumes of 1 mL, 2.5 mL, or 5 mL...necessary MTA, then start production and quality control.  Our packaged on request AAV service uses the ...the same trusted production and quality control processes as our in-stock service: ddPCR, purity testing...blog post on viral vector production and quality control, our AAV Packaged on Request webpage, or our announcement...

  30. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    ...m an introvert!” is not an excuse. Every psych test I’ve ever taken has declared me an introvert, and ...Scientist who coordinates all kit creation and quality control efforts with depositors; Gizela, the lab member...deposits into the repository, performing quality control, and providing technical customer service. Team...easier. Trust Ultimately, your team could have a strong sense of purpose, an excellent leader, and perfectly...
Showing: 391 - 420 of 921 results