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We narrowed to 558 results for: FIE

Showing: 101 - 120 of 558 results

  1. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ...through the expression of a shRNA against p53, a modified reprogramming factor cocktail, and viral protein...trials and remains a safety concern in the clinical field [4]. Three major reprogramming methods are considered...General Hospital with research interests in the fields of gene editing and stem cells. Connect with her...Kotton Four Factors that Differentiate the Stem Cell Field Sign Up to Write for the Addgene Blog! Resources...

  2. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...and using it in mouse embryos to generate gene modified mouse lines. Hopefully this post will help get...HEK293T cells. The fluorescence intensity was classified into four groups (4; brighter than control, 3...base pair sequence surrounding your gRNA. Once amplified, you can insert this region into a pCAG-EGXXFP...matter of transfecting HEK293T cells with your modified pCAG-EGXXFP plasmid along with the individual ...

  3. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ...the applications of optogenetics relevant to your field. Ultimately, your question should dictate your behavioral...retrieval of contextual fear memories [5].  In a modified behavioral paradigm, the authors found that precise...overlooked, aspect of scientific research is that the field benefits from collaboration and the exchange of ...spread of optogenetics throughout the neuroscience field is a great example of this; research benefits when...

  4. Plasmids 101: Control Plasmids

    Type
    Blog Post
    Published
    April 9, 2015, 3:29 p.m.
    ...successfully? Is the shRNA target correct? Is the purified plasmid DNA viable? Or cytotoxic? How and where...over time and from different preparations of the purified test plasmid expressing the independent variable...multiple separate wells (within the same plate) with purified plasmid from the same aliquot/preparation. The...effect of the independent variable because the purified plasmid, cells, and media used in each of the ...

  5. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    Published
    Aug. 15, 2019, 12:30 p.m.
    ... and Valentovich 2014). We routinely clone the purified amplicons upstream and downstream of the desired...included versions with gentamicin-resistance (amplified from TP997). We validated these vectors, which...set of powerful negative selection systems for unmodified Enterobacteriaceae." Nucleic acids research 43.13...system with lambda red recombineering enables simplified chromosomal gene replacement in Escherichia coli...

  6. Plasmids 101: Biotinylation

    Type
    Blog Post
    Published
    Nov. 15, 2018, 1:50 p.m.
    ...aptly named for the chicken, “Avian”, egg it was purified from. Snell later confirmed that avidin was indeed...the culprit in egg white injury by feeding rats purified avidin (Kresege et al., 2004). Avidin is a glycoprotein...Biotinylated proteins or complexes can be easily purified using columns or beads that are coated with avidin...E.coli proteome. Using this knowledge, scientists identified the 13 amino acid sequence required for BirA ...

  7. Evolution of Lab Techniques

    Type
    Blog Post
    Published
    June 21, 2016, 2:30 p.m.
    ...commonly used. In all cases, the result can be quantified by the strength of the signal in comparison to...experimental questions. Real-time PCR detects and quantifies a gene or DNA sequence of interest by recording...in a string of DNA (7). Sanger sequencing uses modified nucleotides, called dideoxynucleotides (ddNTPs... cores to individual labs and even out into the field. Current versions are already commercially available...

  8. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ...antibodies are just the tools you need to push the field forward! Browse the IPI Glypican Collection here...new site-specific retrotransposon elements they identified and characterized (Fell et al., 2025). In retrotransposon... the transposon to new loci in the genome. The modified transposon RNA has untranslated regions (U, blue...biotinylated proteins can then be isolated and identified with tools like mass spectrometry, providing ...

  9. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ...’t look as expected. Once you have cut out and purified your insert and recipient plasmid backbone bands...Your Vector In the ligation step, you mix your purified, cut backbone and insert in a single tube allowing...diagnostic restriction digest of 100-300ng of your purified DNA with the enzymes you used for cloning. Run...more details. Once your complete plasmid has been verified, you’re ready to get experimenting!   Additional...

  10. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...Davidson et al. 2012, Tsiatis et al. 2010). As the field moves toward standardized thresholds for validating...(in vitro) Works across all cell types Must be verified with cell-based method IDLV 1% Cell-based Programmable...since cell fixing HTGTS Not reported Cell-based Identifies translocations Limited by chromatin configuration...to maintain a consistent set of standards as the field seeks to generate reproducible, quality data on ...

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

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...CRISPR/Cas9 technology and rapid improvements in the field, it has recently become possible to introduce DSBs...ribonucleoprotein complex (csRNP, Figure 1). We use in-house purified Cas9 protein, which, in our hands, is both cost-effective...(tag ICC) as a percentage of total cells in the field (DAPI). Using the ICC assay as a direct readout ...recombined the single-stranded donor DNA (V5 tag) were quantified by immunocytochemistry assays. Dewari et al, ...

  12. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ...after landslide, only adding more stones to the field. Fortunately, our bodies have been fighting cancer...tumor; chemokine-aided activation of APCs produces modified dendritic cells (DCs) and macrophages (M1 and ...still many important questions in this exciting field of research. One of the most important: How will...successful, cancer immunotherapy must aspire to be a field of healthcare as dynamic as cancer itself. Many...

  13. CRISPR Cheat Sheet

    Type
    Blog Post
    Published
    Jan. 27, 2021, 2 p.m.
    ... crisp new CRISPR material for you!  The CRISPR field moves fast. That’s why we’re bringing you a brand...functional change in the DNA sequence. Base Editors: Modified versions of CRISPR nucleases that make single ...convert adenosine to inosine. Activate/Repress: Modified versions of CRISPR nucleases that can’t cut DNA...

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

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ... 75,000 pathogenic genetic variants have been identified in humans and cataloged in the ClinVar database...engineered prime editing guide RNA (pegRNA) that both specifies the target site and contains the desired edit ...of RNA stability. Using a CRISPRi screen, they identified a protein that enhances prime editing in mammalian...forward to following new advances in this evolving field! Find prime editing plasmids! Find plasmids from...

  15. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 7, 2017, 1:58 p.m.
    ...after that, in 1994, Prasher’s collaborator Martin Chalfie expressed GFP in exogenous organisms (E. coli, ...and later C. elegans) for the first time. After Chalfie’s work, scientists really started to take notice...Nobel Prize in chemistry was awarded to Shimomura, Chalfie, and Tsien for their work on GFP. This great honor...

  16. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ... Most general-use protocols, unless otherwise specified, were likely developed on the ABI 7000 series....fairly equal, an assumption that is not always verified before use. If you’re using the ΔΔCT method and... analysis methods that pair well with slightly modified CT analysis, so pick one and use it! Tempting ...and Miller KS. 2003. Accurate and statistically verified quantification of relative mRNA abundances using...

  17. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...The field of induced pluripotent stem cells (iPSCs) has been around for 10 years. In that time, scientists...Cesar A., and Gustavo Mostoslavsky. "The evolving field of induced pluripotency: Recent progress and future...Kotton Four Factors that Differentiate the Stem Cell Field Additional Resources on Addgene.org Visit our ... Check out the Yamanaka paper that started the field of iPSCs Find gene-specific plasmids (including ...

  18. Reflections and Looking Ahead

    Type
    Blog Post
    Published
    Feb. 25, 2025, 2:15 p.m.
    ...Request, a service that dramatically expands and simplifies your viral vector options. Almost any AAV transfer..., V., Fotouhi, M., Alqazzaz, M., Southern, K., Alshafie, W., Baker, M. R., Ball, A. R., Callahan, D., ...K. J., Houston, D. W., Kumaran, R., Rego, M., Schofield, C., … Laflamme, C. (2024). A consensus platform...

  20. Embracing Serendipity: A Crucial Element in the PhD Journey

    Type
    Blog Post
    Published
    April 25, 2024, 1:15 p.m.
    ...it is essential to delve deeply into your chosen field, don't shy away from looking beyond its confines...exposes you to discussions beyond your immediate field, fostering a diverse range of perspectives and sparking...approaches Engaging in mindful meditation, taking field trips, and spending time outdoors in nature or visiting...
Showing: 101 - 120 of 558 results