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Showing: 601 - 640 of 1054 results
  1. Tips for a 1st time AAV user (by a Rookie AAV user)

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    Blog Post
    ...Restriction digest screening for ITR deletion is still necessary. To save time and money, I like to miniprep...Production protocol, I take a lazier approach for harvesting my AAV. I skip the PEG precipitation of the HEK293...PEG solution. PEG takes several hours on a heated stir plate to dissolve and requires monitoring so that...autoclaving, which also takes time. Rather than investing all of this effort in preparing reagents, I choose...
  2. 3D Printing Meets CRISPR Cas9

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    Blog Post
    ...impressive, full-color images of proteins. But they are still flat. To truly appreciate the 3D structure of these...models function as thinking tools – eliciting questions like “what does this model representt?" ... "how...creating physical models of proteins that are being investigated in their labs. Soon after we began working with...of the night dreaming about the conformational gymnastics that this protein must undergo as it interrogates...
  3. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

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    Blog Post
    ...important to isolate females from males while they are still virgins. Females begin mating ~6 hours after hatching...the Classical Genetics Simulator website. One interesting thing to know is that males do not recombine ... of Fly Pushing by Ralph J. Greenspan. It’s a fantastic book for newbies on how to work with flies. ... convince him to use Twitter @jonchowphd but he still reads “#” as a “pound symbol”. Additional Resources...
  4. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

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    Blog Post
    ... vector (deposited by David Root of the Broad Institute) has consistently been one of the repository’s...have used the protocol for this plasmid, and if it still being used. After learning of it's continued popularity...is the creation of ZappyLabs. Currently in beta testing, the software aims to be a life science protocol...bring your phone in the lab, though, honestly, this still grosses me out. Maybe it’s just me – I didn’t get...
  5. Tips for Using BLAST to Verify Plasmids

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    Blog Post
    ... the “Align two or more sequences” option to drastically reduce waiting time for search results. Optimizing...with the "Align two or more sequences" option and pasting your expected protein sequence into the Subject...webservices/#pairwise_sequence_alignment_psa Happy BLASTing! Do you have any tips for using BLAST to confirm...plasmid sequencing results or comments on our suggestions? Share your thoughts here to help other labs ...
  6. Antibodies 101: Producing Recombinant Antibodies

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    Blog Post
    ...2020). Harvest and purification At its core, harvesting recombinant antibodies consists of separating...Created with biorender.com   Another method of harvesting uses magnetic beads. In this method, beads are... the efficacy of our antibodies through stress testing. An aliquot of our recombinant antibodies is stored...assays showed that the majority of antibodies are still equally effective compared to those aliquots frozen...
  7. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...editor, the lab started with the Cas13 scaffold, testing a whopping 21 Cas13a orthologs, 15 Cas13b orthologs... the plasmids for the RESCUE RNA editors here! Testing and improving RNA editing from the Cas13b REPAIR...knockdown is more precise than shRNA knockdown, it still displays substantial off-target activity. By mutating...Addgene. East-Seletsky, Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA...
  8. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

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    Blog Post
    ...clicking the ‘Search’ button a new content-aware suggestion dropdown list will let you jump directly to the...types of searches including depositing labs and institutions. For example, search for “Melina Fan” and you... in Parkinson’s disease. Knowing the primary investigator where the plasmid was created can narrow your...plasmids specifically designed for this purpose: Still stumped? Ask an Addgenie The newly launched search...
  9. CRISPR 101: Making a Knock-In Cell Line

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    Blog Post
    .... To kick off a knock-in experiment the first questions are where, which, and what. Identify where in ...like to introduce the edit. Finally, you must stipulate what sequence to introduce, which is done through...HDR edit, you can try one or more of the below suggestions. Disable the other repair mechanisms Mammalian...! Cas12a is a Cas family member which generates sticky ends upon cutting. Cas12a may promote HDR by generating...
  10. Simplify Cloning with in vivo Assembly

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    Blog Post
    ...is currently investigating the molecular biology of synaptic transmission and plasticity.     Javier García-Nafría...co-transformation with linearized vector backbone. DpnI digestion - 1 μl DpnI is added to the PCR mix to remove ... García-Nafría is an investigator scientist at the MRC Laboratory of Molecular Biology. He is interested...
  11. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

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    Blog Post
    ...-wide screens to effectively and efficiently investigate gene function. CRISPR libraries can be used to...labs of David Root and John Doench at the Broad Institute have developed human genome-wide CRISPRko (knock...lowly expressed genes or genes that are more drastically affected by gene activation rather than depletion...resistance in A375 cells. The SAM system (CRISPR/Cas9 Synergistic Activation Mediator) uses an improved activator...
  12. Better Dyeing Through Chemistry & Small Molecule Fluorophores

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    Blog Post
    ...Janelia Research Campus, Howard Hughes Medical Institute. Chemistry is dead, long live chemistry! The discovery.... These impressive advances lead to an obvious question: In this age of GFP and its ilk, why should cell...Janelia Research Campus, Howard Hughes Medical Institute. He would rather be in the lab making new fluorophores...PubMed PMID: 26575238. 5. Crivat, Georgeta, and Justin W. Taraska. "Imaging proteins inside cells with...
  13. Technical Design of a Western Blot

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    Blog Post
    ... It can be so overwhelming that you might just stick with the protocol your labmate handed you — after... contain any denaturing agents. However, you’ll still have to lyse your proteins. For this post, we are...stronger reducing agent than BME and has a less… distinctive smell. BME smells like rotting eggs but has a...However, these systems are expensive, generate plastic waste, and limit the amount of optimization that... may only have access to one or another. Most institutes have moved towards CCD cameras over the past ...
  14. 15 Hot Plasmids from 2017

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    Blog Post
    ...interneurons across species and will undoubtedly constitute a valuable tool for the research community. You...CaMKII and RhoA during induction of structural plasticity in single dendritic spines. The set of CyRFP ...variants to develop five new base editors with distinct PAM sequences, expanding the number of available...xylose-inducible dCas9 paired with a single, unique, and constitutively expressed sgRNA targeting the 5’ end of each...to premature aging and cancer. The Denchi lab investigated an aspect of telomere length regulation called...
  15. CRISPR 101: Any Base Transversion Editors

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    Blog Post
    ...with a different editing window than existing CGBEs. This distinct approach means researchers can now select...the adenine transversion editor AYBEv3. This tool still retained considerable A → G editing activity at ...rates ranging from 4.7% to 30% at tested sites. Still, it opened a new avenue for base editor development...
  16. An Inside Look into Addgene's Dev Team

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    Blog Post
    ...same blind spots, and there wouldn’t be anyone to question these blind spots. Blind spots often result in...collaborates with stakeholders to resolve the questions.  After the code is written, it is reviewed by...these libraries as ingredients to a recipe. It’s unrealistic to plant, grow, and harvest grains every time...news Resources on Addgene.org Check out our job postings Learn more about Addgene's mission ...
  17. Rett Syndrome: A History of Research and Therapeutic Outlooks

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    Blog Post
    ...cardio-respiratory dysfunction, distinctive hand movements, and show autistic-like behaviors.  The role of...animal models results in rapid symptom onset, suggesting that MECP2 is also essential after development...). Beam Therapeutics, co-founded by Harvard Investigators and Addgene depositors David Liu, PhD, Keith...
  18. Supporting Reproducibility with a Connected ELN

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    Blog Post
    ... delve deeper and look specifically at the characteristics and capabilities an ELN needs to facilitate...Goettingen, and several other large research institutions. The following graphic depicts the ways in which...banks and sequencing files is likely stored in institutional and lab servers specifically designed to hold...illustrated in this video, and to data stored on institutional and lab servers. Connectivity to other tools...
  19. Plasmids 101: Broad Host Range Plasmids

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    Blog Post
    ...Understanding the characteristics of natural plasmids and how they function in nature can inform us on...other hosts. For example, the pJD4 plasmid harbors distinct origins (ori1, ori2, ori3) and replication initiation...these plasmids are compatible with.  Table 1: Non-exhaustive list of common broad host range plasmids and ... P. C., & Wong, H. L. (2023). Development of constitutive and IPTG-inducible integron promoter-based expression...
  20. CRISPR-mediated Plant Base Editors

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    Blog Post
    ... PMC5726555. 4. Xie, Kabin, Bastian Minkenberg, and Yinong Yang. "Boosting CRISPR/Cas9 multiplex editing... a single base. If you heard about CRISPR, one question probably comes to mind: can CRISPR be applied ...Ag-biotechnologist in the National Rice Research Institute, Cuttack, Odisha, India. Currently Kutubuddin ...
  21. Plasmids 101: Codon usage bias

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    Blog Post
    ...there are still many more features to consider when designing a full gene. A non-exhaustive list includes...be many possible ways to encode them and what constitutes the “appropriate” codon is not necessarily obvious...PubMed Central PMCID: PMC2970903. 6. Pechmann, Sebastian, and Judith Frydman. "Evolutionary conservation...
  22. AAV Titers: Where Do They Come From And What Do They Mean?

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    Blog Post
    ...methods measure different characteristics of the solution. These characteristics give us information on ... tries to get the other to guess a category by listing off things that fall into that category? Okay, ...do our best to send you a particular lot (if we still have it available). References 1. Fagone, Paolo...
  23. CRISPR Activation: A Practical Guide

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    Blog Post
    ...the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously plastic tool and... University and former researcher at the Wyss Institute for Biologically Inspired Engineering.     Alejandro....2014.09.039 Wong N, Liu W, Wang X (2015) WU-CRISPR: characteristics of functional guide RNAs for the CRISPR/Cas9...
  24. Antibodies 101: Beyond Surface Labeling

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    Blog Post
    ...membrane to enter cells. Depending on the research question and downstream application, one or several of ...are protected. Amine-binding viability dyes can still bind free amines on the cell surface, which yields...As a result, the largely negative live cells can still be easily gated on during the analysis. Conclusion...Harvard Medical School. His projects include investigating negative regulators of T cells and cytokine ...
  25. Transferable Skills Guide: Conflict Resolution

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    Blog Post
    ...customer support, but there are eight other teams who still retain a customer support element amongst their ...manager to explicitly assign the responsibility in question to a specific person or team. What Makes You ... will not be great at resolving conflicts.  Institutional Knowledge: Of the tools I’m mentioning here,...have used your empathy, listening skills, and institutional knowledge to resolve conflicts. Conflict in ...
  26. Antibodies 101: Introduction to Immunofluorescence

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    Blog Post
    ...cellular world. The technique allows you to ask questions like: “Where does my protein of interest live ...corresponding secondary antibodies conjugated to distinct fluorophores to visualize multiple targets at ...Even with all of the resources available, it may still be difficult to find an antibody in which you are...not only answers to your most pressing research questions, but also stunning images (am I the only one with...
  27. Plasmids 101: Expressing shRNAs for RNA Interference

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    Blog Post
    ...something that has fallen out of fashion but is still useful and most importantly, effective. In the research...silencing. RNA interference RNAi is a useful tool to investigate the roles of specific genes or to knock down ...the backbone, shRNAs are most often expressed constitutively, either in lentiviral or retroviral backbones...valuable tool for your research arsenal, and there is still a dedicated field of study for improving these tools...
  28. March for Science

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    Blog Post
    ...of the world functions. Experiments come next, testing the correctness of the hypothesis. The results ...Environmental Protection Agency, and the National Institutes of Health endanger scientists’ ability to uphold...and inclusion through funding and at their own institutions, while many also seek to honor and credit women... funding is required. This is by no means an exhaustive list and every marcher is not necessarily marching...
  29. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...Niemi, a postdoctoral fellow at the Morgridge Institute for Research in Madison, Wisconsin. It is commonly...events on its activation loop, but can be made constitutively active through phosphomimetic mutation of these...phosphoserine) to control for effects of the amino acid substitution. Alternatively, you can express the completely...Ph.D., is a postdoctoral fellow at the Morgridge Institute for Research in Madison, Wisconsin. She is particularly...
  30. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

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    Blog Post
    ...infection, including both common infections and opportunistic infections not usually seen in healthy individuals...infected cells, an idea that became much more realistic with the advent of CRISPR. Using CRISPR/Cas9 SAM...approach, Zhang et al. turned to the CRISPR/Cas9 Synergistic Activation Mediators (SAM) system available from...Young WB, Khalili K, Hu W. CRISPR/gRNA-directed synergistic activation mediator (SAM) induces specific, persistent...
  31. Viral Vectors 101: AAV Serotypes and Tissue Tropism

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    Blog Post
    ...with BioRender.com. AAV vectors have both distinctive capsids and genomic structures. The viral capsid...abbreviated nomenclature (AAV1, AAV2, etc.) to distinguish the capsids from one another. Common serotypes...tissue tropism. AAV5 AAV5 is the most genetically distinct AAV. Though AAV5 is one of many serotypes used...of AAV1 and AAV2, with only six amino acids distinguishing its capsids from those of AAV1 (Issa et al....
  32. Plasmids 101: Dimers and Multimers

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    Blog Post
    ...undigested DNA on a gel as a control when performing a diagnostic digest is a simple but effective method for detecting...assembly from short reads (<300 bp) which cannot distinguish between large repeat sequences within a plasmid...could try linearizing the plasmid via restriction digestion and then run the linearized plasmid on an agarose...independently increase the frequency of base substitution in human cells. J Biol Chem. 2011 Mar 25;286...
  33. Plasmids for Stem Cell Research

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    Collection
    ...Insert Vector Type PI Publication Do you have suggestions for other plasmids that should be added to this...
  34. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

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    Blog Post
    ...the endogenous RecBCD and SbcCD nucleases from digesting linear DNA introduced into the E. coli. Exo: Exo...spontaneous mutations that occur when Red is constitutively expressed. Which recombination system you use...interested in science communication and in vitro diagnostics.     References 1. Costantino N, Court DL. Enhanced...1.16.39. PubMed PMID: 24733238. 6. National Cancer Institute Recombineering Pages. Additional Resources on ...
  35. Plasmids 101: Modular Cloning Applications and Kits

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    Blog Post
    ...constructs (Moore et al., 2016). The variety of constitutive and inducible promoters, linkers, and tags makes...basic biology of and gene regulation in the opportunistic pathogen Acinetobacter baumannii (Brychcy et...delivery. While Fragmid developers were specifically testing CRISPR tools, it should be easy enough for researchers...tools and reagents already in place. MoClo is a fantastic way to save time and money on studies that require...
  36. Live and Let Dye: Self-Labeling Protein Tags

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    Blog Post
    ...you should investigate your options and weigh the potential benefits and costs before investing in one approach...SNAP-OMP25 with Atto590-BG (green), imaged with stimulated emission depletion (STED) nanoscopy. Scale bar...multiple proteins but label each separately before reconstituting, choosing your colors depending on the needs...
  37. Meet Aavery, Blugene's New Virus Friend!

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    Blog Post
    ... name! We thank you for your participation in suggesting names and voting in our poll. Now, we'd like ...didn’t have a name. Last month, we collected suggestions for names and we’ve recently chosen four names... BosLab, a community lab in Cambridge, MA, for hosting our agar art fun!...
  38. Addgene is Stopping Use of X (formerly Twitter)

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    Blog Post
    ...significantly reduced posting on November 20, 2023, and as of today, will stop posting, monitoring, and resharing...with active scientific communities and welcome suggestions in the comments.  X has moved significantly ...
Showing: 601 - 640 of 1054 results