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We narrowed to 671 results for: FIE

Showing: 161 - 180 of 671 results

  1. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    ...professional organization or a scientific society in your field. My leadership training began with the NPA where... within a professional society in my scientific field - ARVO (as mentioned above). I became an official...closer connection with other leaders in my research field should I decide to remain here.  In a time when ... skills can only assist you and make you more qualified for future career opportunities. Becoming involved...

  2. DNA Purification Without a Kit

    Type
    Blog Post
    ...during the extraction procedure. Additionally, purified DNA was free of agarose and could be digested ...contaminants while nucleic acids stick around. Finally, purified total nucleic acids are released by dipping the...directly into a PCR buffer, where DNA or RNA can be amplified. Compared to a commercial magnetic bead-based ...material: PCR product or agarose gel slices Product: purified PCR product or DNA fragments Maybe you’re not ...

  3. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    ...collaboratively discussing recent publications in a specific field(s). Most anyone who has spent any time in academia...the literature, sampling a breadth of work from a field (perhaps even two articles per meeting) would be...people feel confident in their own knowledge of the field. Perhaps interest has simply waned. When attendance... a way to learn about work outside of one's own field in a structured and interactive way. Like any meeting...

  4. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...interactions. The 1980s saw a flurry of discovery in the field of eukaryotic transcriptional activation and cell... And most importantly: Independently validate identified binding partners with other techniques! Minimizing...that your proteins are not post-translationally modified, co-expression of the enzyme responsible for the...Cell. 1985; 43:729-736. PubMed PMID: 3907859. 4. Fields, S., Song, O. A novel genetic system to detect ...

  5. Plasmids 101: Protein tags

    Type
    Blog Post
    ... testing its function. Small ubiquitin-related modifier (SUMO) can help with folding and stabilization...to affect function, His-tagged proteins can be purified using metal-affinity chromatography, usually using...protease around, but it is convenient when used with purified protein since the enzyme cleaves the SUMO tag ...increase its stability activity, can be readily purified using plasmids found in this paper (available ...

  6. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ... ICE software identifies the percentage of genomes that have been successfully modified with insertions...engineering has revolutionized the gene editing field by making experimental workflows considerably easier...edited and unedited (control) populations is PCR-amplified and Sanger sequenced. ICE compares these sequence...

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

    Type
    Blog Post
    ...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...

  8. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ..., targeted genome modifications. Cas9 has been modified to enable researchers to knock out, knock in, ...Prime editors are the second generation of base modifiers available. This system utilizes an RNA template...activator, consists of a dCas9-VP64 fusion and a modified gRNA that is capable of interacting with an additional... to confirm that your target sequence has been modified!  The exact protocol may vary depending on your...

  9. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...searched for this unique sequence of DNA and both identified lymphocyte antigen 6 complex (LY6A) as a receptor...hematopoietic stem cells. Its ligand(s) has not been identified. While LY6A or other Ly6 gene family members ...susceptibility. The Deverman and Wilson Labs independently identified LY6A as a receptor for PHP.B by looking for a...total, they tested 13 strains, with seven strains identified as permissive and six as non-permissive to PHP.B...

  10. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...within the scientific community, as they allow for simplified transformation processes, and can be used by ...equally. Only a handful of plasmids have been identified as broad host range plasmids. The properties ...are not fully understood, but scientists have identified some of the features that confer broad host range...host range replicons, recombinant vectors can be modified further to increase the range of hosts in which...

  11. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...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 ...

  12. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...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...

  13. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...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 ...

  14. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ... 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...

  15. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...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 ...

  16. Evolution of Lab Techniques

    Type
    Blog Post
    ...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...

  17. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...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 ...

  18. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...’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...

  19. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...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 ...

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

    Type
    Blog Post
    ...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, ...
Showing: 161 - 180 of 671 results