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We narrowed to 815 results for: ache

Showing: 161 - 180 of 815 results

  1. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    Published
    April 14, 2022, 1:15 p.m.
    ...existing antibodies. Each cleaved antibody will produce two ~50kD Fab fragments, one for each arm of the antibody...bispecific, meaning that each scFv can target a different antigen; binding of each can be either simultaneous...antibodies can allow you to be flexible in your approach to affinity reagent applications. Whatever affinity...  Rachel Leeson...

  2. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    Published
    Nov. 21, 2019, 1:53 p.m.
    ...depending on the conference you’re attending and who approaches you. You might also have cross-team projects ...picture of virtually every container we used, then for each listed: An “official” name for the container to...pretty Addgene specific) The types or purposes of each given container, when applicable Any other relevant...enough. We also had to enforce and maintain it. Teaching people to use words in a new way is not necessarily...Words are important!”  If you try any of these approaches in your lab or workplace, let me know. If you...you have come upon any other effective approaches for clearing up communications lines in your professional...

  3. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...Picogreen quantification on each and every isolated plasmid sample and normalize each to fall within a 10 fold...make use of a pipeline that takes the raw data from each sample in our pool and individually assembles the...plasmid sequences). As with all things at Addgene, each part of this new process involves a significant ...Confirming tags and fusion proteins. We’ll break down each of these steps below. Aligning the sequence The ... making sure to note the appropriate barcode for each sample. About 24 hours later we unload the FASTQ...

  4. Science communication: 9 Strategies to Get Your Foot in the Door

    Type
    Blog Post
    Published
    Sept. 27, 2017, 1:23 p.m.
    ...looking for outreach volunteers as well. 4) Blog - Hone your writing skills Take the DIY approach and start...engagement - Participate in science festivals and outreach If you are comfortable talking in public, volunteer...Volunteer – Step into science education Do you aim to reach young people? Try networking with your local education... a blog is not only a great form of scientific outreach, it also helps you to hone your writing skills...Center for Communicating Science. If you choose to teach yourself, check out MIT’s OpenCourseware for course...

  5. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...to be less expensive than an ELISA kit. In this approach, viral RNA is first converted to cDNA and then...several different lentiviruses in parallel, this approach should be considered. Similar to the p24 assay... Fluoresence Titering Protocol Another popular approach utilizes the antibiotic resistance genes carried...have integrated into a target cell’s genome. This approach can discriminate multiple integration events, ...underestimating the viral titer. The main drawback of this approach is that it can be quite labor intensive. Target...virus may want to choose more straightforward approaches such as the FACS-based or colony-forming unit...

  6. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ...-colored protein from the jellyfish Cassiopea xamachana (Blanquet and Phelan, 1987). Eight years later...allows photosynthetically active wavelengths to reach zooxanthellae, the coral’s symbiotic dinoflagellates...Chromoproteins also enhance corals’ resistance to mass bleaching, the loss of their symbiotic algae, during heat...cost and resources are limited or for bioart and teaching applications. Find a chromoprotein for your experiment...agar plate. Image: Liljeruhm et al., 2018. For each chromoprotein, the lab determined the color intensities...protein from the mangrove jellyfish Cassiopea xamachana. Marine Biology 94:423–430 . https://doi.org/10.1007...

  7. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ...is to make it detectable. Visualization can be achieved using fluorescent microscopy - even in living ...membrane permeabilized in order for the antibodies to reach their intracellular targets. Therefore, it can be...setup for a different fluorescent signal) and photobleaching i.e. FP stability. One should also be aware...make sure that the selected FPs do not influence each other and thus potentially falsify the signal. As...Pages 2. Cooper GM. 2000. The Cell: A Molecular Approach. 2nd Edition. 3.  CureFFI.org. Cell Biology 04... Susanna Bachle...

  8. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...design hundreds to thousands of different pegRNAs for each desired edit, it simply becomes impossible (or at... efficiency and building prediction tools with machine learning (PRIDICT, PRIDICT2.0).   How to apply...pegRNA efficiencies with pridict.it. This approach leverages the strengths of PRIDICT/PRIDICT2.0 ...chromatin-based prediction tool (ePRIDICT). In summary, machine learning-based prime editing prediction models ...computational biology. His expertise includes applying machine learning techniques to make sense of biology, with...B. van, Krauthammer, M., & Schwank, G. (2024). Machine learning prediction of prime editing efficiency...

  9. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...protocols attached.  Final step: validation  Remember, there is no “one-size-fits-all” protocol! Each library... we have an amplification protocol developed for each individual pooled library; generalized amplification...decide what transformation efficiency you need to achieve. Most pooled library protocols aim for somewhere...weight of your library plasmids. As long as you achieve sufficient coverage from your transformation, any...varies significantly in plasmid size). On plates, each transformed bacteria will be its own colony, and...

  10. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...for in vivo delivery of advanced genome editing machinery such as CRISPR-Cas. Figure 1: Schematic...transduced to target cells. Within target cells, Cas9 machinery is transiently expressed and binds to target DNA...rates of targeted mutagenesis similar to those achieved by isogenic AdVs encoding TALENs targeting the...18% to 65% (Maggio et al., 2014). An alternative approach to NHEJ is to utilize the Homology Directed Repair... Adv vector delivery is often used, the CRISPR approach based on HDR has the advantage of being a potentially...). If sufficient gene transfer and knock-in is achieved, producer cells could produce corrective amounts...

  11. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ...cells, attacking the cellular machinery necessary for division. This approach also kills dividing non-cancerous... of people die from cancer-related complications each year. Despite these grim statistics, new strategies... University of California, San Francisco, is approaching the Ras problem by exploiting the different mutant...the generation of mutant cell lines. Further, for each plasmid in the kit, the transcript most commonly... collection of cancer-related plasmids such that each is linked to a known major pathway. Now, using our...

  12. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ... more to achieve maximum plasmid yield. The optimal growth time should be determined for each plasmid/...number, the number of plasmid copies supported within each bacterial cell in your culture. Copy number is partially...amount of DNA you purify! Additionally, if you've reached your lysate volume limits, the copy number of some...concentration. Add supplements to the media You can achieve higher cell density using a nutrient-rich broth...insufficient gas exchange will prevent your cultures from reaching the desired density. The volume of your flask ...

  13. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    .... More and more CRISPR tools are being published each month, making CRISPR a great choice for your next...expression systems, the typical applications for each, and potential delivery methods.   General considerations...Determine what type of outcome you are trying to achieve: Do you want to permanently knock out or knock-... All of these outcomes can be most effectively achieved with different CRISPR components. Select the appropriate...target cells.  Mammalian CRISPR expression systems Each model system has its own best practices for efficient... summarizes the various components included with each expression system as well as suitable applications...

  14. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...composed of many different gRNA-containing plasmids, each with a different genomic target. The goal of a CRISPR... of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within the genome (see...a heterogeneous population of mutant cells, with each cell or set of cells containing a mutation in a ... plasmids, discerned only by a unique barcode on each plasmid. As such, sequencing CRISPR libraries after...target gene, and maintaining the distribution of each gRNA within the population is key. Loss of representation...gene is actually important for a given phenotype. Each hit from the screen should be independently validated...

  15. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...while not widely known, plasmid assembly can be achieved in vivo using a bacterial recombination pathway...protocol prior to transformation (Figure 1). As this approach requires no special reagents or purification kits..., but have successfully tested DH5a, TOP10, and Mach1. No common lab cloning strain has failed so far....complex assembly of multiple genes. Primer design for each type of modification is shown in Figure 2, and is...site. Encode half the new sequence at the 5’ end of each primer, with an overlap for homologous recombination...in a gene, while adding a GFP-tag (Figure 3). Achieving multiple modifications requires multiple recombination...

  16. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ...Aneesh's research interests include proteomics, machine learning, and visualization for big biology. GPS...: the enhancers! This leads us to a three-step approach for designing an experiment to identify the critical... bit of stochastic magic and Poisson statistics, each sub-population has, on average, one distinct enhancer...know that this matter exists, but traditional approaches to study its function have been prohibitively... (loops) and tertiary structures (globules) to achieve exquisite compaction—on the order of 700 terabytes...collaborative database. Aneesh's research interests span machine learning, proteomics, and user interfaces for math...

  17. Addgene's 20th Anniversary Party

    Type
    Blog Post
    Published
    June 11, 2024, 1:15 p.m.
    ...travel to Addgene's headquarters in Watertown, Massachusetts, for the occasion! Addgenies enjoying the ...makes Addgene — well, Addgene. We're so grateful for each and every one of you!  Throughout the party, we ...majority of the time was spent with Addgenies meeting each other's friends and families, eating delicious food... the party was to celebrate the contributions of each and every Addgenie, from our co-ops to our directors...  Rachel Leeson...

  18. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ...overcome for both of these approaches, such as delivery of the editing machinery to the brain, minimizing...highlights some of the latest breakthrough therapeutic approaches. What is Rett Syndrome? Caused by a mutation ... how to deliver the precise amount of protein to each brain cell to restore physiological levels. Much...Taysha Gene Therapies, founded on Gray’s work, are each expected to advance their respective gene therapies...editase repaired half of the mutant MECP2 RNA in each of three neuronal subtypes of the hippocampus. This...

  19. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ...germline genome editing, but in vivo gene editing approaches have been limited by problems with Cas9 delivery...packaging capacity of AAV (~4.7 kb). While this approach has been proven feasible, it leaves little room...potent Cas9 protein, the Zhang lab took a different approach. They analyzed over 600 Cas9 orthologs and found...effects of SpCas9 and SaCas9, Zhang’s group used an approach called BLESS (direct in situ breaks labeling, ...allowing them to be packaged in AAV.   Split AAV approaches  While finding smaller Cas9 orthologs works for...break a large transgene in two pieces and package each piece into an individual AAV. When a cell is transduced...

  20. How to Write a Scientific Review Article

    Type
    Blog Post
    Published
    Feb. 16, 2017, 3:30 p.m.
    ...studies. We made a table that listed each protein in the class, then for each protein we listed all the studies... ready to write. What actually happened was that each paper taught me a few things and also highlighted...also added citations in real time as I wrote, so each statement was referenced even in the roughest version... data. For example, we summarized how frequently each member of a class of proteins was mutated, as reported...useful because you could easily see how frequently each protein was mutated, you could see how big the studies...
Showing: 161 - 180 of 815 results