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We narrowed to 802 results for: Clu

Showing: 161 - 180 of 802 results

  1. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...depend on target and sample type. Example steps include: permeabilization, antigen retrieval, and sectioning...available, and your experimental goals. Example steps include: counterstaining, mounting sample on slides, and...couple of steps are preliminary steps that are not exclusive to IF, but they can have a significant impact ...you optimize your protocol, always remember to include controls to show that your antibodies are binding...antibodies validate them in a variety of applications, including IF (or IHC or ICC), and will share some of their...

  2. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ... blue light CRY2-CIB1 heterodimerizes or forms clusters under blue light Check out our Optogenetics Guide... One great way to do this is to use CRY2-CIB1 clustering. Fuse either CRY2 or CIB1 to your protein of ...Rab protein vesicle trafficking via CRY2-CIB1 clustering. Image adapted from Aljiboury et al., 2023, under...earlier work on Rab11, the team used optogenetic clustering of Rab8, Rab11, and Rab35 to understand their...attractive strategy for engineering other enzymes. Conclusion We hope this post has illuminated some of the...

  3. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...editing to change the genetic code of plants, including the development of crops with longer shelf life...to engineer the genome of many plant species, including commonly used model organisms like Arabidopsis... Medicago truncatula and several crop species including potato, corn, tomato, wheat, mushroom, and rice... used RNA pol II promoters for Cas9 expression include the ubiquitously expressing cauliflower mosaic ...toolkits for creating plant expression plasmids, including plant CRISPR plasmids from the Yiping Qi lab and...

  4. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...Symptoms of AIDS include rapid weight loss and an increased risk of infection, including both common infections...at increased risk for other chronic diseases, including dementia, gut disorders, nerve damage, and cardiac...could be used to combat this problem, just as ART includes multiple drugs to lower the odds of developing...near the Cas9 cut site, leading Wang et al. to conclude that some Cas9-derived indels may not ablate viral...

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

    Type
    Blog Post
    ...workplace bullying? Threats to professional status – including belittling opinions, public professional humiliation...competence procedures Threats to personal standing – including undermining personal integrity, destructive innuendo..., intimidation, sexual harassment Isolation – including preventing access to opportunities, physical or... withholding necessary information Overwork – including undue pressure, impossible deadlines, unnecessary...unnecessary disruptions Destabilization – including failure to acknowledge good work, allocation of meaningless...

  6. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...technology can be used for numerous applications, including transgenesis, gene-trap screens, and gene editing...manipulations in animal models or cells lines, including stem cells, without leaving behind any accessory...piggyBac™ transposon containing a selection marker is included in a standard homology directed repair template... used for a variety of research applications, including making transgenic cell lines and genetically-modified... using a variety of gene editing technologies including Zinc Fingers, TALENs, and CRISPR-Cas9 nucleases...

  7. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...Wilson and Doudna, 2013). RNAi consists of two non-exclusive gene silencing pathways: transcriptional and post-transcriptional...afflicted by pervasive off-target effects. This conclusion was based upon numerous phenotype discrepancies...importance of using orthogonal methods to confidently conclude the function of a gene of interest. Tools for ...discussed below). Examples of these constructs include pGAPi, which allows easy creation of long hpRNA...experiment are now available on Addgene. These include the construct that allows for easy insertion of...

  8. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...various sources that are difficult to trace. These include the laboratory personnel, the serum, the cell culture...cross-contamination and due to poor lab techniques. These include reusing pipettes for multiple cell lines rather...Contamination! Tissue culture media often have indicators included in the media that turns yellow from lowered pH...the mycoplasma DNA appears in the form of small clusters, apart from the cellular DNA. Fluorescent DNA ... periods and affect the total cell viability. Conclusion To sum up, it is always important to prevent ...

  9. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...high as 1-3%. In light of this finding, we now include guidance on the plasmid page of DIO/FLEx AAV vectors...you intend to use performs in your system. This includes a thorough testing of both the serotype and promoter... is reached you may obtain a false negative or conclude the viral vector is not working.  When using a...off-target expression that can confound your results.  Conclusion Using AAV vectors in your experiments requires...system. It is very rare for a methods section to include all of the variables that need to be accounted ...

  10. MassBioEd Career Exploration Day at Addgene

    Type
    Blog Post
    ... advice for the group of juniors and seniors, including that it’s never too early to start networking!...was the favorite part of their visit for many, including Natalie Reid who said “Seeing the bacteria on ... of programming for scientists!). Careers that include both science and business or programming were new...showed some highlights of his travels from 2015, including a great European photo slideshow from our UK team...

  11. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...2015). This kit offers a broad range of parts, including copy number machinery, chromosomal integration...YTK and adds several additional capabilities, including conditional gene expression, multiplexed CRISPR...chromosomal integration and gene expression, the system includes a toolkit to express recombinant proteins within...kit plus the plant-specific parts you‘ll need, including sub-cellular localization signals, plant markers...engineering. This is a specialized system with parts including site-specific recombinases, regulatory elements...

  12. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ... impact downstream outcomes of an experiment, including the accuracy and efficacy of findings. If you’...peptides and antibodies. Example applications of this include coupling to an antibody, whereby excitation of ...them under experimental conditions. Common issues include photobleaching, non-specific binding, and quenching...biotechnology. Recent examples of advancements include the Janelia Fluor(R) (JF) dyes developed by the...super-resolution applications. Other current advancements include work exploring far-red-emitting dyes in bacterial...

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

    Type
    Blog Post
    ...selection screening. They and their colleagues including Feng Zhang used the GeCKO library to identify ...libraries developed by MIT's Tim Wang and colleagues, including David Sabatini and Eric Lander, comprise 73,171...different gene targets. These pooled libraries include enrichment against kinases, cell cycle proteins...designed to target a smaller set of genes and it includes more guides per gene. The Sabatini/Lander library...

  14. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...building blocks including 14 different fluorescent proteins, 8 constitutive promoters, 2 includible promoters...Characteristics of the pHlash reporter protein include: H+ specific response cytoplasmic retention insensitivity...been used for a variety of experimental purposes including (1) single or multiple, constitutive or inducible...MXS-chaining were used in a variety of experiments including subcellular visualization of 6 different organelles...introduction of many different functional groups, including reactive groups for conjugation reactions, fluorescent... characteristics favorable for innocuous tags including high solubility, limited affinity for other proteins...mammalian cells with Broccoli Vegetable tags, including Spinach, Spinach2, and Broccoli, can benefit your...

  15. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ... lobes, REC and NUC, and their subdomains. NUC includes the HNH and RuvC catalytic domains, as well as...as the CTD (also known as the PI), while REC includes most of the bridge helix. Created with BioRender.com...the cleavage domains, the recognition domains—including the CTD and the entire REC lobe—do not structurally...lobes. The guide RNA in this structure does not include stem loop 3, and guide RNA nucleotides 11-20 are...discoveries on this topic are still being made! Conclusion Finally, we can summarize the process from start...

  16. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    ...replication, and fluorescent proteins that are already included in Snapgene’s extensive database. Snapgene also...of open reading frames, which is often our first clue about where the gene of interest might be located...plasmids are difficult to sequence by NGS. These can include GC-rich and repetitive regions. These regions require... same features annotated in our plasmid maps, including our most popular primers and certain restriction...

  17. Fostering communication in remote collaborations

    Type
    Blog Post
    ...stays on the same page. Ideally, these meetings include all collaborators, from the lab members who are...like Zoom or Google Hangouts is a great way to include researchers from different locations. Having virtual...to your collaborators. This helps everyone feel included in the project and accelerates the pace at which...and morale. Especially when your collaboration includes scientists working in different time zones, it...

  18. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ... and copy number of the plasmid) and typically include a gene that is advantageous for survival, such ...range of applications. Some common plasmid types include cloning plasmids, expression plasmids, gene knock-down...the many things that plasmids can be used to do include: Produce large amounts of a protein so that scientists...the lab. Addgene’s Molecular Biology Reference includes information about molecular cloning, how to choose...

  19. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    ... Now you’re wondering if your favorite gene is included in other CRISPR screens and whether that data ... each screen is captured by BioGRID curators, including the cell line/tissue type, enzyme system, sgRNA...-depth walk through of how to use ORCS:     Include your CRISPR screen data in ORCS  Researchers who...who wish to have their CRISPR screen data included in the BioGRID ORCS CRISPR database are encouraged ...

  20. Using Addgene's New Viral Service for Your Research

    Type
    Blog Post
    ...distributing lentivirus (with many CRISPR tools included among the preps that are currently available) ... AAV preparations have worked well in my lab, including in applications where we have delivered them in...The blog provides tips for working with viruses including advice on how to titer lentiviral preps, troubleshoot...on each viral prep’s specific material page and includes western blots as well as other expression data...
Showing: 161 - 180 of 802 results