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We narrowed to 608 results for: Sis

Showing: 241 - 280 of 608 results

  1. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ...non-dividing cells. The first lentiviral vector system consisted of three plasmids: the packaging, envelope, and...other reporters, and they showed that expression persisted for at least thirty days. Although adeno-associated...integrate into the genome, they could promote oncogenesis by altering local gene expression. Self-inactivating...

  2. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...minigenome, the viral polymerase initiates RNA synthesis at the 3’ end of the minigenome to produce the...These products can be detected by Northern blot analysis. In addition, the polymerase will use the minigenome...mRNA can be detected directly by Northern blot analysis or RT-PCR. Uses of the minigenome system Minigenome...

  3. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...tracrRNA, crRNA guides are only ~42 nt long. Direct synthesis of these crRNAs should be significantly cheaper...rates, determined using sophisticated genome-wide analysis. At many of its computationally predicted off-...at Addgene. 3. Kim, Daesik, et al. Genome-wide analysis reveals specificities of Cpf1 endonucleases in...

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

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...immunoprecipitation (locus-specific ChIP). These methods consist of insertional chromatin immunoprecipitation (iChIP...on development of technologies for biochemical analysis of genome functions such as transcription and ...technologies. The locus-specific ChIP technologies consisting of iChIP and enChIP are recent examples. We also...

  5. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...that a plasmid can transfer to, replicate in, and persist in. Some plasmids have a narrow host range, while...host range recombinant vectors Replicons, which consist of a plasmid’s ori and all of its control elements...scientists can incorporate interchangeable antibiotic resistance cassettes (Prior et al. 2010), inducible promoters...

  6. How to Be an Excellent Trainee

    Type
    Blog Post
    Published
    Sept. 26, 2023, 1:15 p.m.
    ... is the person you report or talk to on a daily basis who will help you get on your feet with techniques...accordingly in case you have questions or need a little assistance at some point during the procedure. Be an active...everything is magically made up by helpful TAs or assistants, and sitting out on the bench waiting for you...

  7. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...concentration, ionic strength and pH, affect binding the lysis buffer may need to be adjusted depending on the ...include protease and phosphatase inhibitors in the lysis buffer, keep samples on ice, and perform the IP ...For mass spectrometry, elute in a urea-containing lysis buffer. Alternatively, the protein can also be eluted...

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

    Type
    Blog Post
    Published
    April 13, 2021, 1:15 p.m.
    ...pTargetF system Our original pCas/pTargetF system consists of one plasmid that expresses the S. pyogenes ...engineering case study of N-acetylglucosamine biosynthesis, we constructed a strain library with 1-11 copies... modified pCas/pTargetF system for CRISPR-Cas9-assisted genome editing in Escherichia coli. Acta Biochimica...

  9. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...CRISPR corner New from the viral service Faster catalysis after yeast-based protein evolution By Joanne ...evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription factor ...CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases, base editors, gRNA backbones...

  10. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    Published
    Feb. 18, 2020, 2:15 p.m.
    ...singletons (reads that only appear once) in our analysis. Many pipelines toss these singletons out, and...stranger - Finding mentors for the meeting Just like analysis pipelines for DNA that group similar sequences...variability was organic, as some people need less assistance than others, but we did have some participants...

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

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...important models for solutions to the coral reef crisis. I like how they are using protocols.io to create...disparate environments so can be used study the genetic basis of evolution in immunology, neurobiology, and more...lab has the capability) with receptor mediated endocytosis. They used this to edited those brown marmorated...

  12. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...They first created a defective chloramphenicol resistance gene (CamR) by introducing a point mutation (... adenine base editor would restore antibiotic resistance. To find such a protein, they created a mutant...any restriction enzyme (RE) recognition site, RE analysis of PCR product could be used to detect success...

  14. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...Properties of FLP Recombinase Evolved by Cycling Mutagenesis.” Nature biotechnology 16(7): 657–62. PubMed ...Deleter Mouse in C57Bl/6 Based on FLPo Recombinase.” Genesis 48(8): 512–20. PubMed PMID: 20506501. 6. Logie,...Synapses Are Repeatedly Assembled and Removed at Persistent Sites In Vivo.” Neuron 89(4): 756–69. PubMed ...

  15. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...proteins within the bacterial genome. From this analysis, they identified 53 potential candidate genes ...within Cas13a are essential for RNA cleavage, consistent with known roles for HEPN domains in other proteins...located at the 3’ end of the spacer sequence and consists of a single A, U, or C base pair. In bacteria,...

  16. Bioinformatics at Addgene

    Type
    Blog Post
    Published
    July 14, 2022, 1:15 p.m.
    ...the accompanying sequencing data collection and analysis eventually outpaced our software, even with continuous...singular purpose (ex. adapter trimming, quality score analysis, k-mer counting, or sequence assembly), but the...would handle plasmid sequences on a 96-well plate basis, but for rapid testing and refinements, we specified...

  17. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...of its parts. Elements such as the antibiotic resistance gene and the origin of replication are necessary...elongation factor 1 alpha Constitutive  Tends to give consistent expression regardless of cell type or physiology...expression, but may vary by cell type. Tends to resist promoter down regulation due to methylation or ...

  18. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...ability to reassemble fragmented genomes is the basis of several split vector approaches developed in ...transgene expression from split vectors has been inconsistent. A greater understanding of basic AAV biology...vector strategies and lead to greater and more consistent transgene expression. References: Allocca M...

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

    Type
    Blog Post
    Published
    Dec. 2, 2019, 2 p.m.
    ...promiscuous biotin ligase (Kim et al 2016). BioID2 consists of a humanized biotin ligase from A. aeolicus,... assemble when in close proximity. Split-BioID consists of a split BirA* molecule that can reassemble ...compared to the standard E. coli BirA* mutant. RapID consists of this mutant BirA* and an RNA component that...

  20. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...s group created Spinach2 using site-directed mutagenesis of Spinach. In doing so, they identified the ... IV, were then identified through microfluidic-assisted in vitro compartmentalisation for their high binding...PMC3852148. Warner, Katherine Deigan, et al. "Structural basis for activity of highly efficient RNA mimics of green...

  21. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...contributed by guest blogger Joachim Goedart, an assistant professor at the Section of Molecular Cytology...conditions needs to be examined on a case-by-case basis. GFP is sensitive to acid The chromophore of GFP...blogger, Joachim Goedhart! Joachim Goedhart is an assistant professor at the Section of Molecular Cytology...

  22. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    Published
    Aug. 23, 2016, 2:30 p.m.
    ... social science to an audience that I thought consisted of all biologists. The social scientists in the...to them with an answer, and do so if you can. 9. Resist the urge to say everything I allow 1 slide for ...topic or story is all you need, if you do it well.  Resist the urge to show all or even a lot of data.  I ...

  23. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    Published
    June 23, 2016, 2:30 p.m.
    ...must be recoded to ‘TGA’ using site-directed mutagenesis. In the C321.ΔA strain, this TGA will be recognized... formation: the last puzzle of aminoacyl‐tRNA synthesis." FEBS letters 462.3 (1999): 302-306. PubMed PMID...Sauerwald, Anselm, et al. "RNA-dependent cysteine biosynthesis in archaea." Science 307.5717 (2005): 1969-1972...

  24. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ...C-terminal HA tag. Selectable and stable: A puromycin resistance gene makes stable cell line creation a breeze...supernatant and selected with puromycin. Puromycin-resistant cells were fixed and labeled with primary antibody...the genome (Figure 4). Figure 4: STITCHR consists of a nicking Cas9 fused to a shortened R2Tocc ...

  25. Finding Your Perfect Job After University

    Type
    Blog Post
    Published
    Jan. 12, 2016, 3:30 p.m.
    ...particularly interesting as I had just completed my thesis on the study of e2Fb, which is a tumour suppressor...but I really struggled with the large scale data analysis, as this is increasingly reliant on programming... to use diagnostic tool for the detection and diagnosis of different types of pancreatic cancer and highlighting...

  26. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...as a control. DNA was analyzed by PAGE. The basis of the Surveyor assay is that Surveyor nuclease ...edit. Using Surveyor nuclease followed by gel electrophoresis, these mismatches are indicated by the presence...Figure 4). 5. In theory you could use densitometric analysis to quantitate the intensity of the bands on your...

  27. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...undesirable side products generated during chemical synthesis of these molecules. To make large amounts of pure...applications, but there are more! Engineering disease resistance through gene stacking, increasing production ...Generation of 8× Nicotiana Benthamiana and 12× Arabidopsis Mutants.” The Plant Journal, vol. 106, no. 1,...

  28. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    Published
    Oct. 24, 2017, 1:54 p.m.
    ...depends on it. Deanna McLafferty is an editorial assistant and activist. Washington, DC, USA 38.9072°N 77.0369...scientists – including global warming skeptics – insisted that more data and research funding are critical...trying turn a few exoplanets into a successful PhD thesis. We use the same atmospheric physics to understand...

  29. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...plasmid vectors and/or rely on cumbersome in vitro synthesis of single guide RNAs (sgRNAs), are time-consuming...reagent preparation in the lab. Figure 3: csRNP-assisted epitope tagging is scalable to 96-well format...., Gogolok S et al. 2017. Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic...

  30. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...number. First you generate four unique kanamycin-resistant plasmids, each containing a different gRNA target... all but one of your gRNAs into spectinomycin-resistant plasmids pX330S-2 to pX330S-(last gRNA number).... Yin, Linlin, et al. “Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs...

  31. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...engages the prime editor protein. The prime editor consists of a Cas9 nickase fused to a reverse transcriptase... domain for optimization. They developed phage-assisted evolution strategies to evolve more effective ... J., Osborn, M. J., & Liu, D. R. (2023). Phage-assisted evolution and protein engineering yield compact...

  33. A Conference By Postdocs For Postdocs: Future of Research

    Type
    Blog Post
    Published
    Sept. 2, 2014, 3:05 p.m.
    ...career landscape and there is insufficient (or no) emphasis on developing transferable skills to enable pursuit...recent discussions of this apparently worsening crisis recently appeared in eLife: "A Fair Deal for PhD...

  34. Microbes: Look, Listen, and Tell

    Type
    Blog Post
    Published
    Aug. 28, 2018, 12:04 p.m.
    ...Saftic) to educational gifs created by the Amoeba Sisters. Here’s one example of how they “bridge the jargon...jargon gap using gifs”: https://amoebasisters.tumblr.com/post/139944465237/lactose-around-we-can-make-...

  35. Science Communication Snapshot: DayCon 2015

    Type
    Blog Post
    Published
    June 17, 2015, 9:04 p.m.
    ...lectures targeting three broad concepts: the genetic basis of disease, cancer, and viral infections. An interactive...science, but they didn’t seem to mind! Attendees consistently mentioned how pleased they were to learn about...

  36. Save Time and Money by Making Your Own Competent Cells

    Type
    Blog Post
    Published
    Jan. 28, 2014, 3:10 p.m.
    ...you're transforming a plasmid with ampicillin resistance.  After a plasmid is introduced to bacteria, ...culture without antibiotics to give the antibiotic resistance gene time to be expressed.  However, you can ...

  38. Supporting Rare Disease Research with BeHeard 2017

    Type
    Blog Post
    Published
    July 20, 2017, 2:30 p.m.
    ...genes as drivers of peritoneal mesothelioma tumorigenesis. They plan to use CRISPR and lentiviral plasmids...peritoneal mesothelioma cells lines and assess their tumorigenesis. These experiments will serve to validate these...

  39. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    Published
    Feb. 20, 2014, 4:57 p.m.
    ...they also went in search of genes involved in resistance to the cancer drug vemurafenib. Enriched libraries... screen for genes involved in bacterial toxin resistance, identifying 4 previously unknown genes.  “With...

  40. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    Published
    June 10, 2025, 1:15 p.m.
    ... evolution took over (with a little mutagenic assistance). Quicker CAST activity allowed certain mutated...evoCAST to the Liu lab’s own PASSIGE (Prime-Editing-Assisted Site-Specific Integrase Gene Editing), which combines...
Showing: 241 - 280 of 608 results