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We narrowed to 767 results for: SCR

Showing: 161 - 180 of 767 results
  1. Antibodies 101: Single Chain Fragment Variables (scFvs)

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    Blog Post
    ... rounds of screening. Created with Biorender.com.   Some phage display libraries screen antibody genes...expression systems. scFvs are also small enough to be screened for with in vitro display methods such as phage...and VH sequences. Phage display also allows for screening scFv generated from the B cell antibody genes ...the ways antibodies are. scFvs are even used to screen for new antibodies. To do this, antibody variable...variable domain sequences are expressed as scFvs and screened with in vitro assays to select for strong binders...
  2. Site Directed Mutagenesis by PCR

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    Blog Post
    ... are isolated from the resulting colonies, and screened for the desired modification. Finally, the positive...final circular plasmid product pictured here. B) Screening of recovered plasmids for the mutation of interest...mutation). Similar results can be obtained with PCR screening using the primers specified by the arrowheads ... vastly facilitates the subsequent process of screening for succesfully mutated clones. Forward and reverse...can also form colonies under these conditions. Screening If a restriction site was introduced (or ablated...ablated), bacterial colonies can be screened by identifying the presence (or absence) of that particular ...
  3. Viral Vectors 101: Systemic Capsids

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    Blog Post
    ... Table 1 gives a brief overview of published descriptions of commonly used or emerging capsids. Selected...2017). AAV-PHP.eB has frequently been used for screening candidate regulatory elements such as enhancers...type-specific promoters or Cre lines. Spatial transcriptomics suggests that AAV-PHP.eB, when paired with ...capsid of choice for future promoter/enhancer screening. However, AAV-PHP.AX packages viral genomes at...target gene expression (EGFP, in the depositing manuscript) across the brain. AAV-BI-28 has been tested ...when using a ubiquitous CAG promoter. Spatial transcriptomics shows that its tropism is relatively more biased... Gradinaru lab. It was selected from a library screened in marmosets. AAV.CAP-Mac effectively transduces...
  4. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

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    Blog Post
    ...from the event can be found below along with a description of the program beneath it. You can jump to the...the different time stamps in the description to watch any sections you’re particularly interested in, ...abbreviated methods section found within a published manuscript. Luckily, many organizations have been working...JoVe, and Bio-protocol all work to provide more descriptive and interactive protocols that make it much easier...would get in-principle acceptance of the final manuscript regardless of whether the ultimate results were...Minisymposium on Reproducibility Booklet - Including descriptions of many organizations promoting reproducibility...
  5. Technique: Probe Phage Genomes for Host Binding Proteins

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    Blog Post
    ...identify possible gene function. In this post, I will describe a useful and cost-effective method that our lab...and probe with live cells The following is a description of a technique published by Simpson, Sacher, ...be cloned into E. coli and subsequent proteins screened for cell-binding abilities (6). This can be accomplished...long-term library storage. After overnight growth, scrape up, pool and extract plasmid DNA directly from ...construction for functional genomic and metagenomic screening." Applied and environmental microbiology 74.5 ...Protein-Protein Interactions Use CRISPR For Genome-Wide Screens Resources on Addgene.org Visit Our Microbiology...
  6. What's New in CRISPR - September 2019

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    Blog Post
    ... fluorescent proteins, fusion proteins, or transcriptional regulators. In this system, Cas9 generates ... Find the CasX plasmids Drug inducible transcriptional activation and genome editing Yu Wang's lab...developed drug inducible CRISPR/Cas9 systems for transcriptional activation and genome editing by fusing Cas9...library with SpdCas9 to perform pooled CRISPRi screens to identify essential genes and phage host factors...metastasis cohorts and a prior mouse metastasis CRISPR screen. The Chen Mouse Tumor Suppressor Gene CRISPR Knockout...
  7. 6 Steps to Submitting a Resume That Gets Seen

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    Blog Post
    ...experience. Most Addgene PhD level job descriptions clearly describe that the roles do not involve bench ...submission of a job application. 3) Read job descriptions carefully Be open minded and be bold. You don...qualifications to apply for a position.  A job description is usually a wish list and the hiring manager... way to a hiring manager. Learn to confidently describe your strengths in your resume and don't be shy...
  8. Antibodies 101: Antibody Engineering and Directed Evolution

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    Blog Post
    ...efficiency! Screening mutant libraries After generating your mutant libraries, you have to screen thousands...generating the library of mutants and how they are screened. When conducting antibody evolution experiments...across large mutant libraries, high-throughput screening techniques such as ELISA and cell surface display... on the surface of cells. This enables direct screening via flow cytometry for improved binding under ... for Protein Engineering: library generation, screening, and affinity maturation. Methods in Molecular...
  9. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

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    Blog Post
    ...sequence prior to an ATG start codon to prevent transcription in cells where the recombinase was not present... transgene into the correct orientation for transcription. Upon recombination one of lox2272 sites will...expression. To test their hypothesis, they randomly scrambled one pair of lox sites and either used the same...transgene (homologous pairs) or used different scrambled sequences to flank the transgenes (non-homologous...amplification. On the left homologous pairs of scrambled cre-recombinase sites lead to greater frequency...leaky expression. On the right non-homologous scrambled recombination sights lead to reduced reversion...
  10. Revamp Your Lab Meetings With Creative Virtual Collaboration

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    Blog Post
    ...make the screen-sharing experience more interactive. The button is on the right of the Share Screen control...control panel on the top of the screen and every participant get to annotate things as they come up in their...their minds. The final screen can also be saved and exported as a picture. I believe that showing something...pivotal in writing publications/grants too. As described by several works (Wuchty et al., 2007, Vicens ...
  11. Choosing the B(right)est Fluorescent Protein: Photostability

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    Blog Post
    ...of Amsterdam. The previous post in this series described a practical approach to selecting a bright fluorescent...literature are performed in different ways. We describe a number of issues with some of the experimental...fluorescence, which, in its simplest form, can be described by a mono-exponential decay (analogous to radioactive...). Consequently, the intensity decay is often described by the t½, which is the time after which half ... al, 2008; Bindels et al, 2017) and cannot be described by a single parameter. Therefore, it is necessary...
  12. Plasmids for Endogenous Gene Tagging in Human Cells

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    Blog Post
    ...or gene copy. The coding sequence for GFP is transcribed and translated along with the gene, producing... last exon of the gene to ensure it would be transcribed and translated. For an N-terminally tagged protein...each cell line. The editing strategy and design described above can be used to create similar donor plasmids...Endogenous gene tagging with fluorescent proteins as described here enables live cell imaging to study cellular...
  13. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ...CRISPR pooled libraries that allow functional screening of thousands of genes in genome-wide experiments...dividing and non-dividing cells. More detailed descriptions of the lentiviral elements and the 2nd and 3rd...regulation, gene of interest expression, knockout, and screening. There are many more useful viral vectors available... lab. Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral...collection can be used for in vivo phenotypic screening of small-molecule libraries and can be combined...
  14. Fluorescent Tagging of Endogenous Genes with SapTrap

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    Blog Post
    ...user wants to carry out a high-throughput genetic screen, it is expensive and time consuming to individually...each insertion. Also, it remains challenging to screen for the genetically modified strains that contain...simultaneously introduce a selection marker for screening modified strains. With SapTrap, the user first...selectable marker (floxed Cbr-unc-119) for easy screening of the insertion event, and a variety of connector...hundreds of genes, generating powerful genetic screening libraries. Additional protein tagging systems ...
  15. Plasmids 101: Knockout/Knock-In Plasmids

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    Blog Post
    ...template with the endogenous gene. Here, we'll describe the techniques and the plasmids used to inactivate... a rare event, so you must select as opposed to screen for cells where recombination has occurred. Neomycin...Conditional knockouts The methods and plasmids described in the preceding sections are simple ways of knocking...knockout, researchers often use the Cre/Lox system described earlier. In this case, you design your targeting... this gene will still be expressed. You'd first screen for this specific recombination outcome using PCR...
  16. CRISPR/Cas9 FAQs Answered!

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    Blog Post
    ...nickase system is based on the Cas9 D10A nickase described in Figure 4 of the Cong et al. (2013) Science ... The human U6 promoter prefers a 'G' at the transcription start site to have high expression, so adding...co-transfect in HR donor. Alternatively, a new system described recently is the double nickase system, with PX335...destroy the BbsI sites, a double digest should discriminate between positive and negative clones. Clones..., how can cells with that mutation be selected/screened? A13: Before starting your experiment, consider...sequencing, for example) on each colony in order to screen for positive ones.  -- OR -- If your HR template...
  17. QC Sequencing Technologies at Addgene

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    Blog Post
    ...developed a separate script for manually inputting known innocuous differences. This script compares the sequence...disregards the specified discrepancies. Any sequences that are a perfect match from this script can then continue...time, but does not leave much flexibility for discrepancies in backbone sequences. Most theoretical sequences...
  18. Hot Plasmids - November 2023

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    Blog Post
    ...in vitro and in vivo. Incorporating reverse transcriptase (RT) variants, PE6a and PE6b are 500–800 bp ...LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID, where two proximity labeling enzymes...recently created a system for recording the transcriptional history of a cell. Lin et al. generated a fusion... HaloTag dyes and showed they could record transcriptional activation events as bands of induced GFP-iPAK4...Analyzing the ticker tape images could recover transcriptional histories, accurate to within 40 minutes. ...
  19. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

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    Blog Post
    ...you can use to study these genes! Groucho, a transcriptional co-repressor The first conference room, that...Addgenies, was named after the protein Groucho, a transcriptional co-repressor important in development. Groucho.... It binds to a specific set of DNA-binding transcription factors to repress the expression of genes primarily...Ish-Horowicz. "The Groucho/TLE/Grg family of transcriptional co-repressors." Genome biology 9.1 (2008): ... Jiang, Jianqiao, and Helen White-Cooper. "Transcriptional activation in Drosophila spermatogenesis involves...
Showing: 161 - 180 of 767 results