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We narrowed to 838 results for: IGH@

Showing: 321 - 360 of 838 results

  1. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    Published
    April 7, 2022, 1:15 p.m.
    ...optimal for experiments requiring high signal to noise ratios, imaging of high frequency glutamate signaling...to create SF-iGluSnFR. This improved variant has higher expression levels in bacteria and produces stronger...properties The improved variants of iGluSnFR3 have higher expression levels and produce stronger fluorescent...Biosensors A Practical Approach to Choosing the B(right)est Fluorescent Protein Visit the Fluorescent Protein...

  2. Trading Spaces - Visiting Scientist Edition

    Type
    Blog Post
    Published
    Aug. 22, 2023, 1 p.m.
    ... be ideal for you. Here, we will chat about who might want to explore this role, how to seek out such ...commitment, planning, and funding – they may not be right or necessary for many scientists! Unless some of...from third parties or your home institute. Discuss rights Do you expect to generate data that will contribute...It is never too early to discuss authorship and rights for anything that could be generated during the...opportunity, but also a really good time! If it sounds right for you, start thinking outside of your current ...

  3. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    Published
    April 4, 2023, 1 p.m.
    ...buck - it grows at higher cell densities than most of its counterparts, yielding higher production power... phaffii in the 1970s with the goal of producing high protein animal feed. Due to increasing prices of...flask volume. What’s even better? K. phaffii is highly efficient at protein secretion, making harvesting...knock-in or knock-out) is essential to your work, you might want to pick a yeast strain best suited to it. Putting...

  4. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ...figure to the right), we identified R-body mutants that extend at both lower and higher pH. We initially...putting the operon under a strong promoter results in high R-body expression  (Schrallhammer et al 2012). This...environmental conditions. More speculatively, R bodies might endow divergent organisms with new functions. For...understanding of how R bodies assemble and function, we might be able to endow cells with new ways to interact...

  5. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    Published
    March 24, 2016, 2:30 p.m.
    ...sides of the break, we determined that Cas9 held tightly to three of the four strands of the cleaved duplex... the PAM-proximal side, consistently supported higher frequencies (up to 60%!) of gene editing than single...have shown that this technique supports extremely high frequencies of genome cutting and, when donor DNA...included in the nucleofection reaction, extremely high frequencies of HDR. The detailed protocol for nucleofection...Central PMCID: PMC4383097. 3. Kim, Sojung, et al. "Highly efficient RNA-guided genome editing in human cells...

  6. Finding and Joining Your Dream Lab

    Type
    Blog Post
    Published
    April 8, 2014, 2:50 p.m.
    ...In a big lab, the advisor might not be around much and your real boss might be a postdoc – this isn't ...competitive style to spur you to succeed or you might enjoy working closely with others to accomplish ...before you take the position. Am I choosing the right mentor? Make sure your advisors will be good role...candidate to ask lab heads, lab members and lab neighbors during the interview process. What if I want...

  7. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ... you to select cells with high GFP expression. These cells have the highest levels of Cas9 and gRNA expression... which method you use, the procedure will vary slightly.   Two Golden Gate options that are available...expression, and thus the highest frequency of genome editing events. Yamamoto Lab Multiplex CRISPR/Cas9.... “Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing.”  Nature Biotechnology...al. “A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and...

  8. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    .... If submitting plasmid(s) using a spreadsheet (right), click the “Download Deposit Spreadsheet” button..., and plasmid description/experimental purpose (right). On the article page, enter the name, type, and...The more sequence data available, the better. We highly encourage you to upload full sequence data whenever...cannot be propagated in DH5α. For plasmids with highly repetitive sequences (which can be prone to recombination...Figure 12: The "Copy" button is located near the top right of each data entry page. Once you have completed...necessary by my institution or other third party rights holders. Once this process is complete, you will...

  9. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    Published
    June 29, 2021, 1:15 p.m.
    ...hybridoma, you can choose one that produces high-affinity/high-avidity antibodies. The same hybridoma is...drive can be identified by sequencing the heavy and light chains. To circumvent any possible genetic drift...  Figure 2: Changes to the heavy chain or light chain genes within a hybridoma impacts the specificity...validating monoclonal antibodies, their cost is higher than that of polyclonal antibodies. For example...

  10. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    Published
    May 3, 2018, 1:48 p.m.
    ...RNA without altering the genome, but Cas13s have higher on-target specificity. New work from Konermann ...package in low-capacity vectors! These small, but mighty type VI-D enzymes are the latest tools in the transcriptome...including Cas13d-specific regulatory proteins and high targeting efficiency, both of which are described...Cas13d deposit here). Konermann et al.: CasRx is a highly potent, specific editor in mammalian cells Not ...characterized Cas13 enzymes, CasRx also displays very high on-target efficiency; where shRNA treatment produced...

  11. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

    Type
    Blog Post
    Published
    Dec. 6, 2018, 1:34 p.m.
    ... Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...here.  Listen to the hot plasmids episode! New, highly efficient RNAi construct for C. elegans gene knockdown...noroviruses can infect millions annually. Because it is highly contagious, it is essential to contain outbreaks...binding experiments. Potential experiments that one might perform using the purified domains include protein...

  12. The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It

    Type
    Blog Post
    Published
    Jan. 30, 2020, 2:15 p.m.
    ...cognitive biases such as motivated reasoning and hindsight bias, we may fool ourselves and our readers into...replications are unavoidable, but can be minimized with high powered research designs. Unanticipated discoveries...become public eventually). It is typically done right before data collection starts, and should include...example, if you hypothesize that growth rates will be higher in treatment versus control, and you think there...of the study, say a fifth time has unpredictably high rates, should be reported as a new discovery that...

  13. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    Published
    Nov. 24, 2020, 2:15 p.m.
    ...is building the software tools for the right people at the right time. To explain a bit further, a software... a strong background of undergraduate research straight to a lab career at a biotech company, I always...operations software for next-gen sequencing (NGS), and high-throughput screening. While some developers had ...about what a software product manager does, you might be wondering what the day to day looks like. Check...

  14. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    Published
    Jan. 21, 2025, 2:15 p.m.
    ...realized that working in the wet lab wasn’t the right path for me. I began exploring alternative career...track and feel supported. Another team member was highly self-motivated and excelled when given the freedom... made similar transitions can provide you with insights, lessons from their experiences, and practical...perspectives and facilitated a team meeting to highlight their complementary strengths. By encouraging ...

  15. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    Published
    Nov. 5, 2019, 1:28 p.m.
    ...sensitivity. Wild-type Cas9 exhibits high on-target activity, but also high off-target activity, which may ...sensitivity is the measure of on-target activity (right). Cas9 nucleases that exhibit enhanced site ...et al., 2018) and found that Sniper-Cas9 showed a high degree of both specificity and sensitivity across...Evolved Cas9 variants with broad PAM compatibility and high DNA specificity." Nature 556.7699 (2018): 57. PubMed...

  16. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...close proximity to the GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The...the AAV capsids (left) and transduction patterns (right).  Both AAV-PHP.eB and the AAV.CAP-B capsids are...mammalian cells. Long sequences traditionally have higher risks of off-target mutations. These plasmids contain...

  17. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...to induce high levels of gene activation for both reporter and endogenous genes. It is highly specific ...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...target only requires a small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the...

  18. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...mixture of products or from incomplete PCR — although higher DNA concentrations are needed for this method to...retain products with single-stranded overhangs (right branch). The mixed PCR and incomplete PCR methods...with two 5′ overhangs, so these approaches require higher concentrations of insert to succeed. 5′ overhangs... showed that a five-piece assembly reaction was highly efficient (~80%) with 40-bp homology regions. Ten-fragment...method also based on homologous recombination. The higher temperature at which Gibson assembly takes place...

  19. Clear and Accessible Writing

    Type
    Blog Post
    Published
    Feb. 15, 2024, 2:15 p.m.
    ... visitors with information about images that a sighted reader would glean from the image itself.  As we...impaired readers receive the same information as sighted readers.    Figure 1: An image from AAV...image, we thought it would be most helpful to highlight the technical details of collecting the purified... passage of supernatant through each layer. The right panel illustrates a gradient tube before and after...

  20. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    Published
    Feb. 28, 2023, 2:15 p.m.
    ...Since its early years, the light microscope has been a powerful tool in almost all disciplines. In biology... cells, has become an established alternative to high-priced commercial devices.     Fig. 2: Workflow...Workflow for open software (left) vs. open hardware (right).    Taking this all together, the essential...open hardware. Upper left panel shows the path of light through a microscope. Lower left panel shows an ...differentiating monocyte with planes and hardware to it. Right panel shows workflow of open hardware.    Conclusion...

  21. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...surprise then that a lot of our subject matter comes straight from you, whether you realize it or not! Many ...what basic information someone new to antibodies might need to successfully start using them at the bench...bench, along with questions we thought they might have as they go. The guide was then assigned to our science...deeper expertise of our antibody team to rely on. High-impact content like the antibody guide always gets...sketch a western blot out quickly." Final design (right). Immediate and immense regret of the graphic designer...

  22. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    Published
    Nov. 10, 2022, 2:15 p.m.
    ...does this actually look like? For a selection, you might put cells into a drug-containing media in which ...scientific question can help determine which is right for you. Screen variations Forward and reverse screens...method you choose to assess traits of interest is highly specific to your scientific question. Things to... assay system include: How specific is it? Is it high throughput? Will it be sensitive enough? No one ...screens Selections sound like a great shortcut, right? Who would ever do a screen when a selection can...

  23. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    Published
    Dec. 21, 2021, 2:15 p.m.
    ...direct approach (left) versus the indirect approach (right). Primary antibodies are represented in blue; secondary... purple; signaling molecules are represented in light green; and proteins are represented in orange. Created...only benefit of secondary antibodies. Finding the right primary antibody for your assay and protein of interest...antibodies are different species and/or isotypes. For highly multiplexed assays, using secondary antibodies ... gel-based method identifies proteins solely by weight and size, relying on the signaling molecule solely...

  24. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    Published
    April 23, 2018, 2 p.m.
    ...Every few months we highlight a subset of the new plasmids in the repository through our hot plasmids...Chemistry for his contributions to super-resolution light microscopy, comes from his recent Nature Communications...difficult to study in living cells using conventional light microscopy due to their small size (<250 nm), and...probes had to target endosomes, be exceptionally bright (which is necessary for effective STED), and be...

  25. Reproducibility for Everyone: Lessons from an Open Science Collaboration

    Type
    Blog Post
    Published
    Feb. 28, 2019, 1:34 p.m.
    ... and international team assembled the right people at the right time. Be inclusive and seek solutions...Teytelman’s vast network and ability to identify the right opportunities, he was able to bring together talented... email the team. Acknowledgements One of the highlights of 2018 for me was to collaborate with this incredibly...

  26. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    Published
    Nov. 21, 2019, 1:53 p.m.
    ...conference you’re attending and who approaches you. You might also have cross-team projects within your own lab... errors in our LIMS tracking system. Though you might not have to communicate with a team of software ...document, I update the training frequently. In hindsight, getting a list of frequently used words like ...everyone on the same page. Others in the meeting might also have the same question and asking can help ..., meetings will be shorter and more productive. Higher Productivity. Taking active measures in improving...

  27. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    Published
    Sept. 8, 2020, 2 p.m.
    ...research. Congratulations to the winners! Below, we highlight their research and how they will use plasmids ...research into CCDS. The brain and muscles have a high metabolic demand that is powered by ATP. A molecule...bicuspid aortic valve at a rate that is 50-100 times higher than women in the general population,” says Dr.... and can lead to cardiac developmental defects. Eight genes from the X chromosome are expressed at significantly...

  28. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    Published
    June 27, 2019, 12:25 p.m.
    ...growth of diverse species. These results further highlight the need for in situ genetic tools for microbiome...Yeast Toolkit by replacing the backbone vector’s high-copy number bacterial ColE1 origin with the RSF1010...microbial population dynamics in the gut can provide insight into how the microbiome responds to infection, ...on the assembly of fluorescent proteins into one bright particle that segregates into one cell during cell...

  29. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ...the entire editing system with a simple LED light. Lighting up your gRNA with fluorescent dyes In a scientific... nucleic acids with N-methyl substitutions are slightly more efficient and less toxic to mammalian cells...less than a minute of exposure to the appropriate light. The photoactivatable counterpart to this system...unable to engage a target sequence. Upon a brief light exposure (several seconds) the gRNA is released ...

  30. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    .... The protein of interest is then eluted using a high imidazole buffer that outcompetes the interaction...protein but not that of the protein of interest. A high imidazole elution buffer disrupts the interaction...concentration of imidazole used in the wash buffer should be high enough to disrupt contaminant-metal ion binding ...therefore more selective. When using systems with higher levels of histidine-containing protein contamination... histidine residues that are present, the more tightly the protein will bind to the metal ion substrate...

  31. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ...Methods to scale up multiplexed CRISPR systems for high-throughput screening are vastly useful for mapping...CombiGEM uses iterative one-pot reactions with straightforward restriction digestion and ligation steps to... by sequencing its set of barcodes. CombiGEM is highly flexible and can accommodate any genetic elements...Central PMCID: PMC4253859. 4. Hart, Traver, et al. "High-Resolution CRISPR Screens Reveal Fitness Genes and...PMC3972032. 10. Wong, Alan SL, et al. "Massively parallel high-order combinatorial genetics in human cells." Nature...

  32. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ..., these problems can be overcome by screening a higher number of potential clones. Golden Gate assembly...constructs with an attB recombination scar encoding eight amino acids, but Golden Gate assembly can be designed...that may not be amenable to "domestication", you might want to consider using an alternative method like... for cloning complicated constructs in a single, high-efficiency step. References: Sequential amplification...one pot, one step, precision cloning method with high throughput capability. Engler C, Kandzia R, Marillonnet...

  33. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...cell lines to the -omics data. The cell lines with higher expression of the target protein, as determined...expression, are often preferred for this approach. A highly selective antibody should be able to detect even...only be able to detect the target protein at the high levels caused by overexpression. Additionally, if...clones The antibody validation process represents a high level of effort and time, which may not be available... & ABIF Consortium. (2024). A guide to selecting high-performing antibodies for Synaptotagmin-1 (Uniprot....15467 Virk, H. S., Rekas, M. Z., Biddle, M. S., Wright, A. K. A., Sousa, J., Weston, C. A., Chachi, L....

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

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...possible by the use of our high-throughput DNA isolation process that yields high-quality isolated DNA samples... on our ability to provide requesting scientists high quality data! Addgene's new in-house NGS process...be familiar with our efforts to ensure access to high quality, ready-to-use viral preps and pooled libraries...Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F (2017) Genome-scale CRISPR-Cas9...

  35. Rabies and Neuronal Tracing

    Type
    Blog Post
    Published
    May 29, 2018, 1:51 p.m.
    ...these high levels of viral replication are toxic. Second, infecting subsequently higher and higher order...transmission occur, spreading the virus through higher and higher order neural connections. While this transsynaptic...

  36. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...of the ABE8s ranged from 1.5x higher for positions A5-A7 to 3.2x higher for positions A3-A4 and A8-A10...synthetic library of TadA sequences to create the eighth generation of adenine base editors, called ABE8... in variants named “ABE8.x”: “8” stands for the eighth round of ABE evolution and the “x” is the placeholder...editing frequencies. For applications that require high DNA editing specificity, for example for potential...

  37. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    Published
    Jan. 9, 2020, 1:45 p.m.
    ...installment of the Early Career Researcher Toolbox will highlight four tools for finding journal articles related...summary of the topic and a chronological list of highly cited papers that contain your search term. When...of the page, as well as the number of citations highly influenced by this paper. Influence is measured...number of publications that heavily cite, or are highly influenced by the author. You can also see who ...

  38. Lab Automation at Addgene

    Type
    Blog Post
    Published
    March 22, 2018, 1:28 p.m.
    ...Updated Apr 6, 2021. If you’re doing high throughput work, it’s likely that you come into the lab wishing...transformation One protocol that has been automated is high throughput transformation. With our ever growing...the deck of the Biomek. The FXP id also able to do high throughput DNA preps and other molecular work if... through this process. With enough agar and the right program, the liquid handler can fill 400 vials in...hand. How can you start automating your lab? You might not be doing enough transformations, minipreps, ...

  39. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    Published
    July 18, 2019, 12:55 p.m.
    ...expression and degradation. These workshops not only highlighted new tools but also included time for questions...presented at the C. elegans meeting. Here I’ll highlight a few toolkits that we have available at Addgene..., and Erik M. Jorgensen. "SapTrap, a toolkit for high-throughput CRISPR/Cas9 gene modification in Caenorhabditis...

  40. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...above, or other. High or Low Copy Choose from: high or low copy. Choose “high copy” if a sufficient...standard cloning strain DH5α. For plasmids with highly repetitive sequences (prone to recombination in...Example of the deposit spreadsheet showing the first eight columns and a pop-up instruction box for the “Purpose...
Showing: 321 - 360 of 838 results