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We narrowed to 846 results for: tat

Showing: 261 - 280 of 846 results

  1. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...within this kit contain a variety of promoters to facilitate gene expression in bacteria, yeast, mammalian...His, Avi, and GFP) as well as protease sites to facilitate protein purification. We hope this kit will allow...important microalgae: intrinsic suicide switch implementation in cyanobacterium Synechocystis sp. PCC 6803...Furthermore, some plasmids contain peptides to facilitate protein folding and/or protease sites to remove...The gene set was carefully chosen to perform statistically relevant benchmarking of cell-free expression...By counting DNA sequence reads, FREQ-Seq can quantitatively determine allele frequencies across timepoints...selective pressures: Miller, Corwin, et al. "Adaptation of Enterococcus faecalis to daptomycin reveals...

  2. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ... introduce naturally occurring cancer-causing mutations into the endogenous allele of the STAG2 tumor ... introducing non-sequence specific frameshift mutations through aberrant double strand break repair, it...knockins, it is not always ideal. Because of these limitations, the AAV approach is best suited for tissue culture...epitope tags, or the introduction of dominant mutations of a gene into the endogenous allele in cultured...template to mouse zygotes and thereby correct a mutation that causes Hemophilia B. Unfortunately, work ...applications that require large gene fusions. The current state-of-the-art in combining the AAV and CRISPR-based...segregation in human cells harboring tumor-derived mutations in STAG2." PLoS genetics 12.2 (2016): e1005865...

  3. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...biotin ligase, BirA*, with a site mutation (R11G). This site mutation destabilizes the ligase, allowing...the E. coli version. It also contains a single mutation (R40G) in a conserved residue in the biotin catalytic...ascorbate peroxidase for proximity labeling A major limitation of APEX was its low sensitivity. Sensitivity ... two mutants, the 35 kD TurboID, which has 15 mutations compared to BirA and the 28kD miniTurbo, which...which has the N-terminus domain deleted and 13 mutations compared to wild type BirA. TurboID and miniTurbo...interactions Split-BioID combines a protein fragment complementation (PCA) technique with proximity labelling to...

  4. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...template to create mutations HDR donors can contain insertable sequences or specific mutations, along with homologous...sequence at the DSB, or it can introduce specific mutations into the damaged DNA if these modifications exist...HDR efficiency for more details on how "Cut-to-Mutation distance" affects editing efficiency. One important... cutting and repairing your target DNA until a mutation is introduced disrupting one or both of these ...sequences. To prevent re-cutting and the resulting mutations after an HDR event, ensure the gRNA or PAM site...off-target effects. A technique that pushes the size limitations of ssODNs is Easi-CRISPR (Quadros et al., 2017...

  5. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    ...purified GFP could be imaged in the green state after excitation at 488 nm, photoconverted with 405 nm light... light, and then captured in the red state after excitation at 561 nm. This imaging process worked well...S65T GFP, and no red color was observed after excitation at 488 nm, indicating that spontaneous conversion...al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP photoconversion One concern with photoconverting...

  6. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...detect proteins in different states. For example, in immunoprecipitation (IP) the antibody interacts with... for protein interaction studies such as immunoprecipitation and pull-down assays.  Westerns are divided... with a protein in its native state while a western blot detects denatured polypeptide chains. The protein...epitopes, differ between a protein in its native state versus the polypeptide chain. Consequently, antibodies... way to gauge protein abundance they provide qualitative measurements. If you require precise quantification...determine protein interactions following immunoprecipitation and are a common readout for subcellular ...

  7. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ...knock-ins of patient mutations which are variants of unknown significance – mutations with unclear effects...with defined, straightforward causes (a specific mutation in a known gene = disease onset) may be best served...accessible, fast, and easy. In cases where the mutational basis of a disease is known – repeat expansion... can aid in generation of multiple edits or implementation of expression systems to accurately mimic a...CRISPR could even be used to generate patient mutations and then test both individual and combinatorial...

  8. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ... H161Y, P163T, I175Q, C176T and N177F mutations). Its excitation maximum is reported as 603 nm, while ...aptamers and MS2-binding fluorescent proteins were the state of the art in RNA imaging in vivo. These new “Riboglow...et al. PLoS One, 2016) and tested a series of mutations. Through this process mCarmine was developed (...several cell types is the lack of a rapid and quantitative editing assay. To help accelerate the expansion...-UGI-mediated activity edits out a frame-shift mutation within the mCherry reporter. Quantification of...

  9. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    ... job asks for experience in “statistical analysis”, mention “statistical analysis” (word by word) in your...general format of a resume and the different presentation styles. Wikipedia does a good job summarizing...handled large data sets and are familiar with statistical analysis. If you have done fluorescent imaging...publishing or patent application processes? How about facilitating a collaboration between research groups? Perhaps...universities make their job postings and salary expectations public, so start by looking at these university...

  10. OCaMP: A New Calcium Indicator for Neural Imaging

    Type
    Blog Post
    ... their excitation wavelength. OCaMP is well-suited for these lasers, with two-photon excitation at 1030...3: One- and two-photon excitation and emission spectra of OCaMP. 1P excitation peak is 545 nm, 1P emission...available in your microscope setup. Figure 1: Excitation (dotted) and emission (solid) spectra for various...emission peak is 565 nm, and 2P excitation peak is 1050 nm. Reproduced from Aggarwal et al. 2025 under a...

  11. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...incorporation of a desired mutation into your plasmid of interest by PCR generates mutated plasmids with no methylation...only the non-mutated and methylated template is destroyed leaving behind a pool of mutated plasmids which...dam-/dcm- strains may have an increased rate of mutation (as these would also be deficient in mis-match...Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org Check ...

  12. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ... that cluttered our old plasmid maps. Clearly Annotated Features, Enzymes, and Primers - The new maps ...SnapGene’s software positions restriction enzyme annotations throughout the map at locations better aligned...map, you also have the ability to to download the static image file (PNG, see above) for your own reference...sequencing results: On this page you can download an annotated GenBank File or a SnapGene file for the sequences...of choice. If the depositor provided a custom annotated GenBank file, this can be found on the main plasmid...hosted here retain all the features shown in the static maps discussed above, but also have interactive...

  13. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...the immune mechanisms of the host. The cause, manifestation, diagnosis, recurrence, and treatment of cancer...molecular targets under research include TIM-3, LAG-3, STAT-3, EGFR, CD20 and CD73 on specific solid as well..., tumor penetration, inadequate concentration, mutation of cell surface targets by tumor cells, are a ...currently the only FDA-approved DC therapy for prostate cancer and is aimed at increasing life expectancy...radiotherapies have caused increased death receptor presentation on tumor cells thus making these cells more ...differ in practically every facet starting from manifestation to their specific interactions with different...the most important: How will cancers evolve and mutate based on the immunotherapeutics we use on them?...

  14. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...light-sensitive protein QuasAr2 and systematically mutated it. Using a robotic picking approach to select ...2015) or ratiometric Bimolecular Fluorescence Complementation (rBiFC, Grefen and Blatt, 2012). The 2in1 Plasmid...vectors with all possible combinations of tag orientation (NN, NC, CN, and CC). The 4 kits also contain...proteins that bind to either the light- and dark- states of the photoswitchable cPYP, which changes in conformation... partners that bind to only the light- or dark- state of the photoswitchable proteins offers a powerful...binders (which bind to either the light- or dark- state of the photoswitchable proteins) fused to mVenus...

  15. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    Blog Post
    ...-invasive gene delivery and optogenetic excitation. Excitation of neurons deeper in the brain may still...makes them light-responsive. When light shines on excitatory channelrhodopsin-expressing neurons, the channelrhodopsins...action potential (Figure 1). This light-induced excitation can take place in a tissue culture dish but can...high-intensity light for activation which requires implantation of fiber-optic cables into the brain. ...still require fiber-optic cannula implantation, but altering ChRgers so they can be activated with tissue-penetrating...

  16. Transferable Skills Guide: Public Speaking

    Type
    Blog Post
    ...cartoon of the protein.) In the middle of my presentation, lost in a sea of data, my mind went completely...taught me an incredibly valuable lesson: Your presentation will be better if you take time when you need...need it, rather than trying to push through a state of panic. Now, if I feel myself getting too nervous...sense. As tempting as option two was in my panicked state, only option one would let me accomplish my goal...often as needed until you get to the end of the presentation, at which point you will have achieved your ...gig. Being able to give a clear, informative presentation, speak at conferences, or even just provide ...

  17. What's New in CRISPR - November 2021

    Type
    Blog Post
    ...dominant negative mutation in MLH1 (PE4 = PE2+MLH1dn and PE5 = PE3+MLH1dn). Silent mutations added near the...the Cas12f scaffold gRNA and making strategic mutations in the DNA binding cavity of Cas12f. The CasMINI...  and base editing, whereas a different set of mutations (the “V3.1” version) works best for genome editing...protein PEG10 specifically binds its own mRNA to facilitate its packaging into virus-like particles. Feng...boosts the efficiency of prime editing to correct mutations in a variety of target genes and cell types. Their...

  18. Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing

    Type
    Blog Post
    ...articles. Then just start reading and following citations through the literature until you have all of the...they mean. Could there be any other possible interpretation of the results? Then think deeper about the...Is the data of sufficient quality (error bars, statistics, clarity in images etc)? Did they perform all...doesn’t mean that there are no mistakes or misinterpretations. After all, the science was performed, written... brim with reprints and papers with scribbled notations in the margins. If you prefer, printing and hand-writing... Department of Biological Sciences at Louisiana State University. She is a fan of photosynthesis and writes...

  19. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...elegans. To study the role of 3’UTRs, the Arizona State University (ASU) Biodesign institute developed a...polyadenylation for all protein coding mRNAs, and annotations extracted from other databases (Mangone et al...Dynamics Database (WDDD) The WDDD aims to acquire quantitative data on all the cell divisions of the C. elegans...methods provides unique opportunities to develop computational methods to better understand animal development...additional SapTrap plasmids used for introducing point mutations that was published in a microPublication (Schwartz... E. M. "SapTrap vectors for introducing point mutations with unc- 119 + selection." MicroPublication 1...

  20. Hot Plasmids - October 2020

    Type
    Blog Post
    ...you'll find in this post: Tools for labeling excitatory inhibitory synaptic proteins Fluorescent protein...corner New from the viral service Labeling excitatory and inhibitory synaptic proteins with FingRs ...developed AAV vectors to selectively visualize excitatory and inhibitory synapses in the brain.  ...scaffolding proteins PSD95 and gephyrin to label excitatory and inhibitory synapses, respectively. Once expressed... fluorescent proteins targeting inhibitory or excitatory synapses or that are Cre-inducible, researchers... development, regeneration, and certain disease states. However, it’s difficult to observe the cell cycle...
Showing: 261 - 280 of 846 results