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We narrowed to 1,106 results for: Ide

Showing: 281 - 300 of 1106 results

  1. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    ...programs provide support for academic career development, it's often left to students to identify and build... design variables need consideration? Enrich your knowledge and skills outside of the lab Effective verbal...you build a network outside of the academic world Participating in activities outside of the lab expands...and government. This great resource allows the undecided to learn about a variety of science professions...on their skills and interests. Once scientists identify potential career pathways, education and mastering...mastering new skill sets must occur by doing work outside of the lab. Extracurricular activities don't have...tool and technique development. However, careers outside of the lab highlight “transferable” skills such...

  2. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...specificity is dictated by a DNA guide sequence rather than an RNA guide as with Cas9. Third, it can be ...with an ssDNA guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo...Cas9/gRNA complexes or with guide RNA- and Cas9-expression plasmids. Cas9 guide RNA delivery does however...sharing the stage with much-lauded CRISPR-Cas9 is DNA-guided endonuclease NgAgo! We'll discuss how NgAgo is ...have their own pros and cons (2). The NgAgo DNA guide is inexpensive and can be 5’-phosphorylated in house...modified from a company. However, unlike an RNA guide, it cannot be produced from a plasmid in cells. ...Try NgAgo for Yourself On the other hand, the widespread success of CRISPR-Cas9 across multiple model ...

  3. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...Yiqun, et al. "CRISPR Cas9-guided chromatin immunoprecipitation identifies miR483 as an epigenetic modulator...system, consisting of dCas9 and targeting gRNA, to identify molecular interactions at this site (Fujita and...Subsequent mass spectroscopy studies allowed for the identification of proteins associated with the IRF-1 locus...different fields of study. In addition to the identification of DNA binding proteins, this method can characterize...sensitive and specific and allowed the Xu lab to identify protein and nucleic acid binding partners at numerous...various biological fields. In 2015, the Fujii Lab identified non-coding RNAs associated with telomeres by ... malignancies. Additionally, the Tapscott Lab identified regulators of DUX4, a transcriptional regulator...

  4. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...skills guide Resources on Addgene.org Learn more about Careers at Addgene Watch out career videos  ...training. Similarly, there are several factors to consider when selecting a postdoc mentor and lab. These...Depending on the position, you may also want some evidence of teaching experience. How do you get there... having sufficient data to write a manuscript. Consider questions like: What figures are missing? For ...explore your system more, you will have a better idea of how to leverage it to ask new and interesting... grad school, you searched for and eventually identified a postdoctoral mentor. A good mentor typically...typically writes the grants that fund your research, guides the research directions of the lab, gives you constructive...

  5. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...to a target polypeptide. Multimerization in the SunTag system occurs via antibody-peptide labeling; specifically...they righlty deserve, but, in this post, we'll provide a small sampling of the many amazing new plasmid...-2xNLS-VP64 and dCas9(N)-FRB-NES). This system provides users greater temporal control over CRISPR/Cas9...successful in regulating gene transcription using DNA-guided optogenetic tools; however, targeting the necessary...overcome many of the limitations intrinsic to DNA-guided systems such as TAL effectors or Zinc Finger Nucleases...Nucleases, the Gersbach lab has modified the RNA-guided CRISPR-Cas9 system to create a tool that is quick...flexibility of the LACE technology makes this system widely accessible for many potential applications. Polstein...

  6. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...go wrong. We hope that we have provided you with a few ideas to consider if you do run into a roadblock... faced with a roadblock. In this post, we will provide an overview of some of the common challenges associated...such events, newer lentiviral packaging systems divide the viral elements among multiple plasmids. Second...packaging systems please see Addgene’s Lentiviral Guide. While the third generation packaging systems offer...experiments a bit more challenging. With these considerations in mind, when planning an experiment users ...transgene, and downstream applications should all be considered when choosing the correct system. Recently, ...transfer plasmids with small inserts (such as CRISPR guides) while a single 72h harvest is best for larger ...

  7. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...digest, PPIs are identified via mass spec analysis. Biotinylated proteins are considered possible interaction...distinct functions. Many existing methods for identifying PPIs, such as affinity purification mass spectrometry...spectrometry (AP-MS), lack the ability to specifically identify proteins that interact with a particular protein...the validation of a binary PPI as well as the identification of additional interacting factors. Problems...detect weak and/or transient PPIs. Many PPI identification techniques rely on protein interactors to stay... AP-MS. This requirement often prevents the identification of weak and/or transient interacting proteins...potential interactors with a bait protein but the flip side is that it’s difficult to know which proteins interact...

  8. Which Fluorescence Microscopy Technique is Best for Me?

    Type
    Blog Post
    ...microscopy techniques   Fluorescent Widefield Microscopy Fluorescent Widefield Microscopy is the most common...to solve two drawbacks of widefield and confocal microscopy. First, widefield and confocal microscopy project... visual system relies primarily on contrast to identify individual objects and perceive the world around...When performed correctly, fluorescence microscopy provides a high contrast image in which a bright signal...contrast, color contrast, to an image and thereby provide molecular or structural specificity to the viewer...microscope objective evenly illuminating the entire (wide) field of view. Fluorescent light traveling back...one time, limiting photobleaching. Similar to widefield, the entire field of view is excited at one time...

  9. Great Results Start with Great Standard Curves

    Type
    Blog Post
    ...Here, we will provide an overview of how to create and use a standard curve and provide some general considerations... considerations for scientists planning to use them in their assays.  What is a standard curve? Scientists...serial dilution of a control protein, incubated alongside an unknown sample with the BCA reagent. The absorbance... a known control sample, called a standard. The ideal standard curve has at least five dilutions, with... the data points generated by replicate values provides useful information about the accuracy of the curve... must be included every time the assay is run, ideally with freshly prepared buffer. This is because standard...users aim for an R2 above 0.95.    Additional considerations When running assays that rely on a standard...

  10. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...signal-to-noise ratios in two-photon imaging, making it ideal for video recordings of large populations of synapses...screened a large library of iGluSnFR3 mutants, identifying the two  best-performing variants as iGluSnFR4f...vector that contains everything needed to replicate inside a host cell but with space to insert your genetic...and strong expression after inserting EGFP and provide step-by-step protocols for viral generation and...retrotransposons for DNA editing By Emily P. Bentley For RNA-guided genome editing, think beyond Cas9 with STITCHR:...new site-specific retrotransposon elements they identified and characterized (Fell et al., 2025). In retrotransposon...that moving the RNA untranslated regions (UTRs) outside of the homology elements allowed for scarless insertion...

  11. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    ...newer lab members. You must get an idea of what to expect and decide if it is what is best for you. Some...development? To get a better idea of whether or not a lab head is a good role model, consider questions like: How...publication record show evidence of industry collaboration? If you are considering a career at a small, ...focused on choosing a lab, but almost all of these guidelines can apply to any workplace or job. It amazes ...sections below. Mentoring relationships can help identify your priorities and needs, but sometimes it just...talk to as many lab alumni as you can or schedule video/calls if they are not local. Alumni meetings can...Former lab members can give you a fuller picture. Consider talking to people in the lab across the hall or...

  12. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...the plot that divides it into four, using the location of the cell populations as a guide  (Figure 3.) ...machine that everything inside the gate is one population and everything outside the gate is a different... It can be used to measure protein expression, identify rare cells, or even sort out single cells through... to use increasingly more stringent markers to identify your specific cell population of interest. The...reads/interprets the signals, the axis is instead divided into positive and negative signal at a point on....      Figure 3: It's often easier to identify different populations by imagining (or drawing...roughly group the cells by size. This is done using side scatter (SSC) and forward scatter (FSC), which measure...

  13. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    ...experiment - there are a number of outside factors that need to be considered as well. In this post, we’ll ...overnight, so mark the vial, add the incident to your records, and consider the following questions as you ...ll touch on antibody storage, buffer considerations, and give just the lightest of nods towards conjugates... and illustrious lab career, you will likely accidentally leave your antibody out on the bench overnight...1: Blugene fading into darkness because they accidentally left their antibodies out. It's okay, Blugene...could affect the reaction. For instance, sodium azide, a common antimicrobial agent added to buffers, ...reaction. Many commercial antibodies come with sodium azide already included in the buffer, which will be noted...

  14. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ...common, besides being one of the five senses? GPCRs or G-protein-coupled receptors (GPCRs)! Aside from these...molecule ligands such as clozapine-N-oxide (CNO). DREADDs are ideal for chemogenetics as they are relatively...quick guide to chemogenetics Resources on Addgene.org Check out Addgene's chemogenetics guide Find chemogenetics...with heterotrimeric G proteins on the intracellular side of the cell membrane. Heterotrimeric G proteins ...Gα), beta (Gβ), and gamma (Gγ). G proteins are divided into four subtypes, based on their Gα subunit: ...that activate many GPCRs are still unknown. To identify some of these unknown ligands and their GPCR binding...Roth lab showed that this platform accurately identified 120 known GPCR targets demonstrating its potential...

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

    Type
    Blog Post
    ...transcriptase. The Cas9 nickase is guided to the DNA target site by the pegRNA — a guide RNA that also encodes the... 75,000 pathogenic genetic variants have been identified in humans and cataloged in the ClinVar database...developed in David Liu’s lab in 2019, has added considerably more precision and flexibility to the CRISPR...does it work? First, an engineered prime editing guide RNA (pegRNA) that both specifies the target site...editors are preferred. For instance, if target nucleotides are positioned within the canonical base editing...HDR) stimulated by double strand breaks has been widely used to generate precise changes. However, the ...optimized for each application, and the efficacy varies widely — it still almost always results in higher ratios...

  16. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...to an oligonucleotide depending on your downstream application. Biotin labelled oligonucleotides can be...attached to a photoreactive arylazide group. Under strong visible light, the arylazide group becomes reactive...in the small intestine from the microflora that reside there. Biotin is a cofactor that in its active ...Avidin is a glycoprotein that consists of four identical subunits that can each bind one molecule of biotin...macromolecules it is added to. Biotin also has an exposed side chain that can be easily manipulated to make reactive... biology and biotechnology purposes.  Once a polypeptide is biotinylated (more on this below) it can be...multiple biotin binding sites found on avidin are also ideal for signal amplification during these experiments...

  17. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...Experimental guidelines Primer Design As a rule of thumb, 11 bp of complementary sequence on either side of the...PCR process, you need a high-fidelity polymerase. There are many high fidelty polymerases on the market;...technique that can be used to introduce specific nucleotide substitutions (or deletions) in a tailored manner...detects primer multimerization (purple) which is evident by an increase in the size of the lower band (portion...the plasmid of interest during the PCR reaction. Ideally, your primers should be free of palindromic and...Methylation deficient bacterial strains can be identified by the dam13(-) mutation – you’ll want to steer... exonuclease activity (increases amplification fidelity), and no 5’->3’ exonuclease activity (which could...

  18. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ...there, then start identifying the resources needed to accomplish that goal. 2. Identify the tools in that...past year to get an idea of how much you can expect to use in the future. 3. Identify the skills needed ... clients focus on their experimental needs and consider a budget as a resource checklist. A budget is ...Fluorescence Scopes, Flowcytometers, etc.). After identifying all the major equipment, categorize all of your... needed to do your science. This will help you identify the talent pool that you’ll need to tap for your...materials, and resources that you’ve thoroughly identified. Do online searches, and seek advice from your...salaries in Kansas versus salaries in NYC will vary widely. Therefore, start up awards should reflect salaries...

  19. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ...yellow-light sensitive chloride ion pumps, activation of which results in efflux of chloride ions, subsequent...the injections must be near identical for all of these surgeries. Identify coordinates through a rodent... if you are not familiar with it, check out the video below). That’s not to say that the technique doesn...methodological and experimental limitations and considerations? I recently spent two months in the lab of ... at NIDA and some of the practicalities and considerations you may not find in the materials and methods...use optogenetics The most important question to consider is what exactly do you want to use optogenetics...are also available. Picking your opsin  You must consider whether you wish to stimulate or inhibit neuronal...

  20. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...addgene.org CRISPR pooled libraries Guide to pooled libraries CRISPR guide AAV guide ...expression. Episomes are circular pieces of DNA that reside outside a host cell’s genome. In this state, AAV cargo... for each experiment. Additional considerations One of the downsides to using AAV is the limited packaging...valuable part of the molecular biology toolbox to identify new target genes for drug discovery or to understand...effort. The versatility of Cas effectors allows for a wide variety of screens, including interference, activation...rely on lentiviral systems, a fact supported by the wide array of lentiviral-based CRISPR libraries available...selection). Next-generation sequencing (NGS) to identify genes of interest. Figure 1: Overview ...
Showing: 281 - 300 of 1106 results