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Showing: 241 - 260 of 731 results
  1. Plasmids 101: Biotinylation

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    Blog Post
    ... are essential for processes such as gluconeogenesis, lipogenesis, amino acid metabolism and energy transduction...Kresege et al., 2004). Avidin is a glycoprotein that consists of four identical subunits that can each bind ...non-selectively label DNA and RNA. Photobiotin acetate consists of a biotin molecule attached to a photoreactive...
  2. CRISPR 101: Validating Your Genome Edit

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    Blog Post
    ...of WT and edited cell lines. Then decomposition analysis is performed yielding a graph of edits in the ... Fig. 3: Restriction enzyme screening for resistivity to digestion, indicating targeted edits. ...solely for restriction enzyme sensitivity or resistivity, and thus be a marker for edited cells. As with...Software such as CRISPResso can help with the data analysis. If NGS is not an option, you can still detect...
  3. Treating Muscular Dystrophy with CRISPR Gene Editing

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    Blog Post
    ...off-target editing. In order for the therapy to persist over time, stem cell editing would be highly desirable...improved, with vastly reduced inflammation and necrosis compared to unedited animals. In grip strength...approach, increasing the possibility that edits will persist long-term. In accordance with this result, Nelson...CRISPR Could Be Used to Treat Cataracts and Cystic Fibrosis Read How Homology Directed Repair is Used to Edit...
  4. Plasmids 101: Repressible Promoters

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    Blog Post
    ...systems. Binary systems are genetic tools that consist of two parts, both of which must be present for...allows them to be used together for complex genetic analysis in Drosophila or for precise control over multiple...Desplan. “Power tools for gene expression and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55...transgene expression, lineage tracing, and mosaic analysis.” Cell 141(3) (2010): 536-48. PubMed PMID: 20434990...
  5. How to Design Your gRNA for CRISPR Genome Editing

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    Blog Post
    ..., it would be unwise to make conclusions on the basis of the activity of a single gRNA, and thus diversity...on careful experimental design, execution, and analysis. Many thanks to our guest blogger John Doench....2017.97 Hanna RE, Doench JG (2020) Design and analysis of CRISPR–Cas experiments. Nat Biotechnol 38:813...https://doi.org/10.1038/s41576-018-0059-1 Veres A, Gosis BS, Ding Q, Collins R, Ragavendran A, Brand H, Erdin...
  6. Quick Guide to All Things Lentivirus

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    Blog Post
    ...the Moloney Murine leukemia virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing...generation. The 1st generation of lentiviral vectors consisted of three plasmids encoding 1) the lentiviral vector...Efficient gene transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human... from one batch to another. For experimental consistency it’s a good idea to purify and titer your virus...
  7. Transferable Skills Guide: Teamwork

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    Blog Post
    ...of a senior thesis), I knew early on that I wanted to work with a partner. WPI’s emphasis on teamwork ...decision about something? Fix a problem? Test a hypothesis? Whether you officially call it a “project team... risky shots at the goal rather than going for assists that are more likely to lead to scores. That player...
  8. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
    ...post was contributed by Jordan Ward who is an assistant professor at the University of California, Santa...elegans relied on selective markers such as drug resistance (Chen et al., 2013), fluorescence (GFP; Tzur ...Thank you to our guest blogger! Jordan Ward is an assistant professor at the University of California, Santa...on gene regulation in nematode molting and spermatogenesis. His group also builds tools to improve genome...
  9. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    .... Like other ABEs, the editor they were using consisted of Cas9 fused to an engineered adenosine deaminase...tool for predicting CGBE editing outcomes and assisting sgRNA selection is available at http://www.sunlab.fun...A., Yang, D., Mok, B., Replogle, J. M., Xu, A., Sisley, T. A., Weissman, J. S., Adamson, B., & Liu, D....developed using CRISPRi screens, target-library analysis, and machine learning. Nature Biotechnology, 39...
  10. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

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    Blog Post
    ...a prestigious institution: This criterion is consistently under scrutiny. USCIS wants you to prove that... is very important in this case.  You can still assist your PI in the peer review process, but remember...love of science and technology, Rachel is proud to assist companies and foreign nationals working in the ...dedicated to the practice of US immigration law and assists individuals, researchers, entrepreneurs, companies...
  11. Editor's Choice, August 2016

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    Blog Post
    ...optimize and troubleshoot the system. Site Directed Mutagenesis by PCR & Single Base Editing with CRISPR For ...technology may make some uses of Site Directed Mutagenesis by PCR obsolete! Honorable mentions Of course... in chronological order here: Site Directed Mutagenesis by PCR Google Forums Round Up: First Impressions...
  12. Who Gives a Tweet? 9 Facts About Scientists on Twitter

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    Blog Post
    ...this article: “A systematic identification and analysis of scientists on Twitter.” This paper addresses..., or are friends, are included in this cluster analysis. The closer two dots are to each other, the more... C. R. (2017). A systematic identification and analysis of scientists on Twitter. Plos One,12(4). doi:...
  13. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ... than point modifications. CRISPR base editing persists through multiple cell divisions, indicating that...specificity. In this first, the Liu lab used phage-assisted evolution selection systems to generate ABE8e(... a similar approach. Dali Li’s lab’s A&C-BEmax consists of a fusion of both cytidine and adenosine deaminases... E., Doudna, J. A., & Liu, D. R. (2020). Phage-assisted evolution of an adenine base editor with improved...
  14. Sensing Neuronal Dopamine

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    Blog Post
    ...dopamine. Methods for measuring Dopamine In vivo microdialysis The gold standard method of measuring DA and...neurotransmitters in the brain has been in vivo microdialysis of extracellular fluid (Darvesh et al., 2011...References Darvesh, Altaf S., et al. "In vivo brain microdialysis: advances in neuropsychopharmacology and drug...
  15. FastCloning: A Newer, Simpler Cloning Technique

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    ... these techniques and is a multi-step process consisting of digestion, purification, ligation, and transformation... 1. Celie, Patrick HN, Annabel HA Parret, and Anastassis Perrakis. "Recombinant cloning strategies for...Plasmid Cloning by PCR Perform Site Directed Mutagenesis by PCR Catch up on Your Plasmid Background with...
  16. Sleeping Beauty Awakens for Genome Engineering

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    Blog Post
    ...social and regulatory reasons. 1.) The transposon consists of a mirrored set of inverted repeats (green) ...1) an SB transposase, the enzyme required for catalysis of transposition; and 2) a transposon containing...determined by transposition assays of antibiotic-resistance genes conducted in HeLa cells). In their Nature...
  17. Single-cell tracking of lineage and identity with CellTag

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    Blog Post
    ...labels cells with heritable barcodes. Barcodes consist of an 8bp variable region and a 6 bp fixed region... a sample of cells was collected for scRNA-seq analysis and the remaining cells were maintained in culture...detailed CellTag protocol on protocols.io, code and analysis tutorials on GitHub, and a CellTag lineage data...
  18. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

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    Blog Post
    ...was then screened through successive rounds of mutagenesis for the brightest teal fluorescent proteins that...scientists from Florida State University, they used a mutagenesis approach and found that specific amino acid changes...protein localized to their expected locations. An analysis of mWasabi gave similar results. We hope this ...
  19. CRISPR Challenges: Standardization and Homology Directed Repair

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    ...much more about cellular DNA repair pathways. An emphasis on cellular repair post-CRISPR cleavage When I...Ribonucleoprotein (RNP) Delivery The RNP delivery method consistently leads to high on-target editing and low off-...strategies activate different HDR sub-pathways like synthesis-dependent strand annealing (SDSA) and single-strand...
  20. In Vivo Biotinylation of Bacterial Fusion Proteins

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    ...GST-glutathione. Biotin-based immobilization for kinetic analysis Proteolytic removal of the fusion protein could...could provide a more relevant kinetic analysis, but the trick is to utilize a second immobilization method... purification strategy outlined here provides a basis to determine binding kinetics using a multitude ...
Showing: 241 - 260 of 731 results