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We narrowed to 749 results for: Tro;

Showing: 331 - 360 of 749 results

  1. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    Published
    June 27, 2018, 1:16 p.m.
    ...PubMed PMID:29109157. Nanobody toolkit to monitor retrograde transport at the cell surface Article contributed...developed a set of functionalized nanobodies to study retrograde transport from the cell surface. Their toolkit...treated with biotin–phenol and H2O2, APEX2 labels electron-rich amino acid side chains (e.g. Tyrosine) with...biotin-labeled proteins can them be analyzed by mass spectrometry allowing a researcher to specifically identify...

  2. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ... promoters are upstream or 5’ of the gene they control. Promoters can be organism or tissue type-specific...different levels of expression. For example, CAG is a strong promoter that drives very high, ubiquitous expression...recombinase, are used to spatially and temporally control transgene expression. Lox sites are directional...availability of Cre, lox sites lets researchers control the spatial and temporal activation of a transgene...

  3. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ...gene-editing protein - a DNA-cleaving Argonaute from Natronobacterium gregoryi, or NgAgo for short. Addgene has ..., showing that this protein could cleave DNA in vitro, directed by phosphorylated DNA guides. Unfortunately...RNA guides must be expressed from a plasmid or in vitro transcribed, and changes in gRNA secondary structure... Han. “DNA-guided genome editing using the Natronobacterium gregoryi Argonaute.” Nat Biotechnol. Epub ...

  4. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    Published
    Dec. 13, 2018, 1:14 p.m.
    ...increased yields in adenoviral (Coroadinha, 2006) and retroviral production systems (Shen, 2011). To our knowledge...also improved yields of the AAV2 derivative, rAAV2-retro, but had no effect on AAV5 yields. We further show...preparation. Through sugar detection assays, in vitro transduction, and silver stain approaches we show...Effect of osmotic pressure on the production of retroviral vectors: Enhancement in vector stability." Biotechnology...

  5. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    Published
    Oct. 27, 2016, 2:30 p.m.
    ...chemically or electro-competent E. coli - excess salt causes arcing during electroporation which would ...sequenced for potential positive clones. Once you introduce your 3’-end “A” overhang insert, the magic of ...would cause the electroporation to fail). When incubating at room temperature, it is not recommended that...

  6. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    Published
    Dec. 7, 2015, 6 p.m.
    ... 10. Plasmids 101: Control Plasmids Need help thinking about the types of controls you’ll need for your...provides researchers with many different ways to control gene expression. Get the full scoop from Addgenie...Addgenie Chari Cortez guides us through some common controls and why you might need them. Happy Reading!   ...

  7. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    Published
    Aug. 23, 2022, 1:15 p.m.
    ...abstracts comparing the mortality rates between two controversial health issues. One group read the text from ...design or by their preconceived ideas about the controversial medical issues. Participants gave significantly...methodologically flawed, and biased. As a result, the control abstract comparing two fictional medical issues...and education have more polarized beliefs on controversial science topics. Proceedings of the National ...

  8. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    Published
    July 27, 2017, 1:11 p.m.
    ...Xin hypothesized that she could make enzymes controllable by tethering them to Dronpa, a fluorescent protein...their ability to be precisely spatio-temporally controlled as a key advantage over other inducible enzyme... sugars (including lactose), and a lac operon controlled GFP. When the enzyme breaks down the HMO of interest...Photosactivatable Fluorescent Proteins Use Light to Control CRISPR Genome Editing Choose the Brightest Fluorescent...

  9. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    Published
    April 18, 2019, 12:53 p.m.
    ...pairs from intergenic regions, intronic regions, and a pericentromeric region. All pairs were easily visualized...CRISPRainbow. Both tools were targeted to the FBN3 intronic repeat which contains 22 copies of the gRNA target...locations of the subtelomeric region of the pericentromeric region (PR1), the p-arm (T1), and the q-arm...

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

    Type
    Blog Post
    Published
    Dec. 6, 2016, 3:30 p.m.
    ...The gRNAs targeting EGFP can be used as positive controls for gene editing or gene activation, while the...What you don't need to do is spend your time troubleshooting reagent production, because that's a distraction...from your primary goal. Getting these quality-controlled reagents from Addgene can greatly expedite a ...Research: A Primer for Beginners 5 Tips for Troubleshooting Viral Transductions Additional Resources on...

  11. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ...later, it was shown that putting the operon under a strong promoter results in high R-body expression  (Schrallhammer...bodies could also perform as actuators in microelectromechanical systems (MEMS) devices. They could be functionalized...of armor to protect their host from predation (Petroni et al 2000). With a better understanding of how... Plasmids from This Publication at Addgene 4. Petroni, G., Spring, S., Schleifer, K.-H., Verni, F. & ...

  12. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ...digest as depicted in Lane 2. Controlling methylation Finally, you can control methylation by altering your...only the non-mutated and methylated template is destroyed leaving behind a pool of mutated plasmids which... with the Latest Cloning Techniques Learn to Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis...

  13. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    Published
    Jan. 6, 2017, 3:56 p.m.
    ...unique editing tasks, classic Cas9 nuclease, which introduces double stranded DNA breaks, can be expressed ...Gαi-coupled) DREADD fused to mCherry, all under the control of the human synapsin promoter. Purpose of this...mCherry can be used to induce neuronal silencing in vitro and in vivo (Krashes et al., 2011). The various ...Rumpel S. Analysis of transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, ...

  14. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    Published
    Dec. 21, 2021, 2:15 p.m.
    ...antibodies. The introduction of antibody fragments, such as nanobodies, has introduced new options for...Antibodies 101: Introduction to Immunofluorescence Antibodies 101: ELISAs Antibodies 101: Introduction to Antibodies...

  15. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    Published
    July 14, 2021, 1:15 p.m.
    ...antibody, you’ll want to use the same positive control across experiments to be aware of signal strength...you’ll want to be sure to choose the appropriate controls to be sure the staining you are seeing is real...real. It’s important to include a negative control using serum from the same species your polyclonal antibody...antibody was generated in or a polyclonal isotype control, to account for nonspecific binding. Now that you...

  16. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    Published
    May 11, 2021, 1:15 p.m.
    ...localization. Surprisingly our observations at the electron microscope showed us that, in monkey neurons, ...of the haemagglutinin of the influenza virus, introduced to the field of fusion proteins more than thirty... of interest the membrane.   Figure 1: Electron microscope images showing ultrastructural localization...protein of interest, an alternative could be bicistronic expression of the fluorescent protein with the...

  17. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...the insert. Gateway® technology (developed by InvitrogenTM) is essentially a more modern version of this...teachline.ls.huji.ac.il/72682/tutorials/gateway/introduction.html demonstrating the principal behind the “swapping...CcdB-resistant strain is ccdB Survival™ from InvitrogenTM. Using either DB3.1 or ccdB SurvivalTM, one ...PubMed PMID: 1324324. 6. Tam, JE. and Kline, BC. “Control of the ccd operon in plasmid F.” J Bacteriol. 1989...

  18. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ...AAV (AAV-pgk-Cre) has been used to successfully introduce Cre into cells of interest. What if I need two...of using loxP sites is the inability to tightly control which loxP sites recombine if more than two are...allowing for two separate recombination events to be controlled in parallel. Have you used the Cre-lox system...PubMed Central PMCID: PMC49604. 4. Independent control of immunoglobulin switch recombination at individual...

  19. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ...chemogenetics? Chemogenetics relies on genetically introducing engineered receptors that are only activated ...Expression Using chemogenetics requires successfully introducing receptors to target cells. Targeted and specific...mitigated by using low doses and running appropriate controls (Jendryka et al., 2019), they have resulted in...channel, the PSAM acts as a gating mechanism to control ion flux. The system allows you to directly manipulate...

  20. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 3, 2022, 1:15 p.m.
    ...live cells.  Controls  When designing experiments, it’s important to include proper controls to assess the...quantitative readout of the interaction in vivo or in vitro, and can be used for measuring protein expression... p53 tetramerization in response to DNA damage controls the p53 response. Gaglia G, Lahav G. Mol Syst ...

  21. Plasmids 101: Degron Tags

    Type
    Blog Post
    Published
    March 14, 2024, 1:15 p.m.
    ...advantages degron tags can offer over other protein control methods and compare the commonly available types...etc. Firstly, they are reversible mechanisms of control. Protein degradation can be induced by the addition... Tsien RY, Lin MZ. Tunable and reversible drug control of protein production via a self-excising degron...  Susanna Stroik...

  22. Of Myc and Men

    Type
    Blog Post
    Published
    Jan. 19, 2023, 2:15 p.m.
    ...of interest to better study them in vivo and in vitro, especially in the mid-late 20th century with the...reticulum localization, Munro and Pelham casually introduced the Myc-tag (Munro, et al. 1986; Munro and Pelham... and the Myc-tag today Since Munro and Pelham introduced the Myc tag to the world, it has become one of...this antibody works and allow users to better troubleshoot and design experiments. It turns out that even...

  23. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ... proteins. The Seeger lab developed a robust in-vitro protocol that enables rapid selection of synthetic...sybodies that hypothetically contain ones that bind strongly to your target protein. The Seeger lab designed...selection step involves using a commercial kit for in-vitro translation of the mRNA libraries. Each mRNA molecule...enrichment step for identifying sybodies that bind strongly and specifically to the protein of interest. The...

  24. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    Published
    Jan. 10, 2017, 3:37 p.m.
    ...can be exploited to either inactivate a gene or introduce a desired genetic alteration. Listen to Our Podcast...necessary gRNAs, but it would not be possible to control which gRNA becomes complexed with which variant...represents a straightforward means of exerting tight control over Cas9 nuclease activity while eliminating the...guides targeting one set of genetic loci can be introduced together with truncated gRNAs targeting a different...

  25. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    Published
    May 22, 2015, 4:58 p.m.
    ...experiments attempting to build gene drives in tropical mosquito vectors of diseases such as malaria and...then, using multiple confinement strategies is strongly advised. The effects are multiplicative, and there.... Site-specific selfish genes as tools for the control and genetic engineering of natural populations....Fitz-Earle, M., Holm, D. G. & Suzuki, D. T. Genetic Control of Insect Populations: I. Cage Studies of Chromosome...

  26. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...and would avoid genomic off-targets or indels introduced through non-homologous end joining (NHEJ). Designing...recapitulate or rescue known pathogenic alleles, or introduce a premature stop codon to render an RNA nonfunctional...also adjusted the structure of the guide RNA by introducing a C opposite the target A to be edited. This ...off-target edits around the targeted nucleotide. Introduction of guanine-mismatches in the guide across from...

  27. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    Published
    March 20, 2014, 3:04 p.m.
    ...for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work/life balance....possible to bring all aspects of our lives under control? Balance is a myth and should not even be the real... don’t have children, so my life will be under control.” Think again. Maybe you’ll need to care for an...to do most tasks – try to resist the impulse to control by insisting things always be done your way. Be...

  28. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    Published
    May 21, 2024, 1:30 p.m.
    ...reproducible assay. That information can make you a much stronger candidate!  Additional uses The cover letter is...job with a lot of file management that asks for strong organizational skills. You might think, “I’d be...letter The opening paragraph is the place to introduce yourself and your background. It’s a great place...and is important for the job]. Notice that the intros above allow you to highlight what specific skills...

  29. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    Published
    July 13, 2023, 1:15 p.m.
    ...protocols therefore differ from library to library. We strongly recommend following the protocols provided by ...a low multiplicity of infection (MOI). MOI is controlled by viral titer, so you’ll need to measure your...Addgene uses a physical titer (via ddPCR) to quality control our viral preps, we recommend determining the functional...transduction protocol you can follow and a troubleshooting blog should you run into any issues.  Keep ...

  30. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ...and selection for targets can be done largely in vitro; Easy and reproducible commercial synthesis (those..., Akhtar and Marty, Jean L. “Aptamer based electrochemical sensors for emerging environmental pollutants...systems using immobilized aptamers.” Biosens Bioelectron. 2011 May 15;26(9):3725-36. PubMed PMID: 21419619...FRET in Your Experiments Learn more about GFP Controlling Protein Activity with GFP Resources at Addgene.org...
Showing: 331 - 360 of 749 results