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We narrowed to 809 results for: nin

Showing: 451 - 500 of 809 results

  1. FPbase: A new community-editable fluorescent protein database

    Type
    Blog Post
    Published
    May 16, 2018, 1 p.m.
    ...posts by Joachim Goedhart on the Addgene blog – outlining various important considerations when choosing...contributions (using simple web forms) while also maintaining a relatively strict database design (schema)....inaccurate data submissions, so FPbase includes a versioning system that allows users to see when a given ...

  2. DAP arrays for multiplex gene editing

    Type
    Blog Post
    Published
    July 7, 2022, 1:15 p.m.
    ...potential in vivo applications or multiplex CRISPR screening.   Using tRNAs Luckily, a fun feature of tRNA...given tRNA (derived from human or plant source), by tuning the length of 5’ leader sequences of a given tRNA...engineering of crops or allow for sophisticated CRISPR screenings that would perturb the unprecedented number of...

  3. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    Published
    Aug. 1, 2017, 1:15 p.m.
    ...used TRICK to examine translation of Drosophila patterning gene oskar. Early in development, the osk-TRICK...with the red reporter mRNA. Yan et al. used harringtonine, a translation inhibitor that stalls ribosomes...sequences into the coding sequence and 3’ UTR. Before beginning experiments, it’s important to verify that these...

  4. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    Published
    July 21, 2017, 12:48 p.m.
    ... specific tissue type, you mate your fly line containing YFG under the control of the UAS with a fly line...often very easy to get GAL4 UAS experiments up and running with fly lines that are already available, sometimes...references and web links from my first post and start designing your first fly experiment! Many thanks to our...

  5. Addgene Depositors Get More Citations

    Type
    Blog Post
    Published
    April 20, 2017, 2:30 p.m.
    ...publications from these journals into two groups: those containing plasmids that the authors deposited with Addgene... other reasons for the correlation. However, determining this exactly is difficult. For example, it is... papers in the same journals! So, is all this remaining difference caused by depositing the related plasmids...

  6. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    Published
    Oct. 19, 2023, 1:15 p.m.
    ...Replacing the D’ region of an ITR with a sequence containing transcription factor binding sites can increase...expression at varying capacities. AAV vectors containing the AAV2 ITRs can be cross-packaged with capsids...PMID: 31914802. Savy, A., Dickx, Y., Nauwynck, L., Bonnin, D., Merten, O.-W., & Galibert, L. (2017). Impact...

  7. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    Published
    Dec. 10, 2019, 1:58 p.m.
    ... a only a single gRNA per cell. The viral DNA containing the gRNA then integrates into the genome. Thus...are poorly transduced by lentivirus, and thus screening with available CRISPR libraries was not possible...retroviral mouse CRISPR pooled library can be used for screening of cells that are refractory to lentiviral transduction...

  8. 5 Great Apps for Lab Life

    Type
    Blog Post
    Published
    Sept. 22, 2016, 2:30 p.m.
    ...mass spectrometry app. I digress. When I started learning proper cell culture technique, I tried to find...my lab we do a lot of genetic construction and cloning experiments. Not all of our lab enzymes are made...be beneficial for new researchers or students beginning to learn about biochemistry. The least this app...

  9. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    Published
    Aug. 28, 2014, 3:34 p.m.
    ...cleave the DNA molecules at specific sites and 2) running the reaction on an agarose gel to determine the...the gel will ensure you keep smaller bands from running off; however, you may need to run the gel for longer...such as how to set up your digests and pouring/running a DNA gel.  More Plasmid Eductional Resources:...

  10. Advice for Scientists Starting a Lab

    Type
    Blog Post
    Published
    April 29, 2014, 1:43 p.m.
    ... your academic lab, download Science Careers' "Running Your Lab" booklet. What has been the most surprising...with us every day and in the pub with us in the evening, wanting to know our latest results as soon as ...Resources for Early Career Scientists Science Careers: Running Your Lab (pdf) Advice for Choosing a Research Project...

  11. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    Published
    Feb. 21, 2019, 1:15 p.m.
    ...forward genetics harnesses this at a large scale by screening diverse libraries of genetic variants. Several...enzyme. AID, however, only changes cytidines or guanines to the other three bases, which limits the level...bacteria. Its modular nature provides versatility in defining the mutational rate and editing window of the ...

  12. Q&A with Karen Katz and Carl Paratore, the Newest Members of the Addgene Board of Directors

    Type
    Blog Post
    Published
    Feb. 19, 2019, 1:36 p.m.
    ...reached out and asked if I would be interested in joining the audit committee about five years ago. I was...of the office? Katz: I have a puppy and so I am learning how to train a dog with my daughter. It’s super... was associated with Hale & Dorr and Crowell & Morning. Karen was a founding member of Mass Medical Angels...

  14. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ...are commonly used for Golden Gate cloning or for CRISPR gRNA cloning. With this more functional display...display, you can start thinking about your next cloning experiment early on. Beyond these simple but powerful...

  15. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    Published
    June 1, 2017, 2:30 p.m.
    ... versus eukaryote, background fluorescence). Determining practical brightness When a dish of mammalian...cells is transiently transfected with a plasmid containing a fluorescent protein (fusion) there is huge ... under realistic experimental conditions. By determining the brightness for a set of fluorescent proteins...

  16. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    Published
    Jan. 6, 2017, 3:56 p.m.
    ...Sanjana et al., 2014). It is a dual-vector system, meaning it is typically used in conjunction with a second...(Gi)-mCherry from this virus is cre-dependent, meaning that once injected, the DREADD will only be expressed... vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014 Aug;11(8):783-4. PubMed PMID...

  17. Viral Vectors 101: Biosensors

    Type
    Blog Post
    Published
    July 6, 2023, 1 p.m.
    ... pair on actinin will produce different signals based on changes to the structure of actinin in low vs...associated with specific proteins of interest such as actinin. When tension is high vs. low in the cell the donor...

  18. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...are commonly packaged in 400-500 bp cassettes containing the RNA pol III promoter, gRNA and pol III terminator...gRNA multiplexing, Xie et al. constructed PTGs containing 8 gRNAs targeting 4 MAPK genes, again using rice... gRNA constructs with PTGs. In PTG constructs containing two gRNAs, they found 100% indel frequency at...

  19. Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur

    Type
    Blog Post
    Published
    July 10, 2014, 7:17 p.m.
    ...school and turn it into a biotech company. Transitioning an idea from academia to a startup Mike Koeris...competing in business plan competitions. After much learning, the focus of the application was changed, but...million. The biggest challenge during this time was learning to communicate effectively to non-scientific audiences...

  20. Will You Be My Mentor? Finding and Asking for Mentoring Support

    Type
    Blog Post
    Published
    Feb. 4, 2014, 3:50 p.m.
    ... “Learning is not attained by chance, it must be sought for with ardor and attended to with diligence...lookout for friends or colleagues that have the training or skills you want to learn. Mentors can be senior...asking them to have coffee. This is much less threatening! If they say no, don’t worry about it. It is ...

  21. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    Published
    Dec. 10, 2013, 3:13 p.m.
    ...sequence? Musunuru: The kit includes 832 plasmids containing prefabricated pieces that allow you to build ...sequence in the genome in rapid fashion. The two remaining plasmids in the kit have the rest of the protein...takes two to three days from the moment you are designing a TALEN on paper to a finished plasmid. With our...

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

    Type
    Blog Post
    Published
    Feb. 28, 2019, 1:34 p.m.
    ...this pivotal moment that Teytelman realized that running interactive workshops to train researchers on tools...expanding this team, leading monthly calls, and planning next steps. April Clyburne-Sherin had already ...there, allowing others to have the resources for running their own workshops: April Clyburne-Sherin’s and...

  23. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    Published
    April 14, 2022, 1:15 p.m.
    ... important to know all your options when planning and running experiments. You never know what problems...in an ~50kD protein. Minibodies are bispecific, meaning that each scFv can target a different antigen; ...

  24. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ... power (Shaner et al, 2008). In confocal laser scanning microscopy, fluorescent proteins are excited with...photon-excitation, selective plane illumination, TIRF or spinning disk confocal use entirely different excitation...rates can be summarized as follows: Instead of determining a t½ of a fluorescent protein in a water/oil ...

  25. Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages

    Type
    Blog Post
    Published
    Jan. 11, 2017, 3:30 p.m.
    ...particularly interested in whether institutions planning to raise salaries prior to the injunction are ...injunction (see below). Of the institutions who were planning to raise salaries before the injunction, we have... contacted by a number of postdocs who are now planning unionization at their institutions. One clear ...

  26. Developing Lab Management Software for Biology

    Type
    Blog Post
    Published
    Aug. 25, 2016, 2:30 p.m.
    ...management software is not a requisite for a functioning lab, but it is for a scaleable lab. When you ...mirroring the lab procedures, not artificially constraining them). In the second example we have a separate...-and-true tools means less time getting up and running and more time working on interesting biotechnology...

  27. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ...and many loxP-containing alleles have also been constructed. Additionally, Cre-containing adenovirus (Ad-Cre...only in specified cells or at specified times. Combining this with some of the loxP methods described above...

  28. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...intracellular Ca2+ levels than the original version containing CRY2 without the extra peptide. CRY2clust should...Barrick lab has deposited the pDCAF3-IS5 plasmid containing a synthetic operon that enables the degradation...enzyme required for conversion of xanthine into guanine, and without xanthine supplied in the media, these...

  29. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ...fluorescence from other regions of the cell. By combining vectors expressing different fluorescent proteins...can be used to determine cell cycle length by examining the ratio of blue to red cells. Cells undergoing...Addgene The Cas Hybrid for Multiplexed Editing and screening Applications (CHyMErA) Knockout Libraries express...

  30. How to Prepare for an Industry Interview

    Type
    Blog Post
    Published
    July 9, 2020, 1:15 p.m.
    ... working in the pharmaceuticals industry after earning my Master’s in 2016 and then working four more ...to get enough sleep the night before. Have your morning coffee, if that’s part of your daily routine. Stay...s Science Career Guide Check out Addgene’s job openings  ...

  31. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    Published
    March 12, 2020, 1:15 p.m.
    ...expression in the retina.  Four approaches for designing synthetic promoters  To build a library of synthetic...marker (CatCH-GFP). The lab deposited AAV plasmids containing the active promoters at Addgene. Performance ...expression occurred in one type of cell, with the remaining expression occurring in only Müller glia cells...

  33. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ...photosensitive Cre, Opto-Cre-Vvd, by splitting and joining the protein regions with Vivid (Vvd) photodimers... protein to date By Justin Ng High-throughput screening methods, such as fluorescence-activated cell sorting...s lab developed a new form of FACS-based cell screening, Single-cell Phenotypic Observation and Tagging...

  34. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...underlying genetic causes of some human conditions, opening up new avenues for therapeutics that treat disease...immunological reconstitution with a normally functioning T cell repertoire (1). In this example, it was...Ken Y., et al. "Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous...

  35. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...are used as it eliminates the need for antibody staining. Check Out Our In-Depth Fluoresence Titering Protocol...sorting based on expression of a surface marker, subcloning, expansion, and confirmation. This procedure ... How Lentivirus is Used in CRISPR Genome-Wide Screening Learn How Lentivirus Has Been Used for Lineage...

  36. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    Published
    Oct. 22, 2014, 1:57 p.m.
    ...content. Here are some of my tips to organizing and running a memorable panel discussion. Check out Joanne'...where the questions are obvious, the answers uninteresting, the panel members all say the same thing and...will thank you. Own it. For more great tips on running panel discussions see Guidelines for Great Panels...

  37. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...and gRNAs. Using lentiviral delivery of the two remaining CAPTURE 2.0 components eliminates the need to ...identified and the level of on-target enrichment. 2. Determining the spatial and hierarchical organization of ...Variant for Your Next Experiment Learn More About Designing gRNAs Against Your Favorite CREs Here Additional...

  39. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ...Fluorescent Proteins (FP) available for microscopy, by cloning your protein of interest into a vector encoding...gene names in the figure below to find plasmids containing that particular gene. ...colocalization experiments in a scientific and meaningful way it is important to choose the appropriate...

  40. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...protein fusion. This one protein method simplifies cloning and makes your experimental setup less cumbersome...studies of animal development where events like the patterning of nascent tissues and the formation of limbs... process. How have you used FP timers? Are you planning on using an FP timer in a new and creative way...

  41. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    Published
    Aug. 3, 2023, 1:15 p.m.
    ... unstained sample for each tissue type you are running in your panel if you wish to use the autofluorescence...indistinguishable. This tool is of particular value when designing a panel.     Figure 3: Similarity index...if markers are co-expressed in your cells when designing your panel. If you have two highly overlapping...

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

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ... of DNA through error-prone non-homologous end joining (NHEJ). In targeted DNA knock-in, donor DNA is ...CRISPR-Cas delivery utilized the Non Homologous End Joining (NHEJ) DNA repair mechanism to knock out genes ...https://doi.org/10.1038/nmeth.3075 Lukashev AN, Zamyatnin AA Jr (2016) Viral vectors for gene therapy: Current...

  43. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ...soon developed that allowed researchers to create stunning, interactive images of their proteins. Since the... known to say something silly like “See there, arginine 53 really is right there, at the end of helix ...description of how effective this model was in explaining CRISPR technology to 4th graders, check out Jennifer...

  44. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    Published
    Feb. 6, 2020, 3 p.m.
    ...continue such discussions. Beyond the excitement of learning about new cutting edge science, researchers of...the most important is letting people know it’s happening. Aim for 3-4 weeks advance notice for the first...presenter, figure by figure) if the primary purpose is training graduate students in critical reading. If the ...

  45. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...depend on what downstream applications you are planning, cost, and the amount of protein you need to produce...popular choice for this. Chromatography columns containing appropriate immobilized ligands are incubated...incubated with cell culture supernatant containing antibodies. The antibodies bind the immobilized ligands and...

  46. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...retroviruses are RNA-containing viruses  (Robinson et al., 1965)! You may be beginning to realize that the...Lander, M. R., Pugh, W. E., & Teich, N. (1971). Noninfectious ARK mouse embryo cell lines in which each cell...

  47. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...post, most bacteria have poor non-homologous end joining machinery, so a CRISPR-induced double-stranded ...phagemid delivery systems. Phagemids are plasmids containing a phage origin of replication that can be packaged...using phage-derived vectors to carry CRISPR, we’re turning the tables on the bacteria. Future research will...

  48. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...control elements for each would be relatively large. Cloning the viral genes in this plasmid would be more difficult...is important that all of these plasmids (those containing NP, VP35, VP30, L, and the minigenome) are efficiently...ways: transfecting with an additional plasmid containing the T7 RNA polymerase gene or using cells that...

  49. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    ...barcodes can be purchased from many companies such as Corning. Tubes can even have specific barcodes etched or...barcode reader and an Excel spreadsheet. Start by scanning the sample barcodes into the cells in the same...corresponding Excel sheet.3. Be specific  - When assigning a physical location to your samples, include as...

  50. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ...Victoria and expressed in any organism while still maintaining fluorescence. The protein structure, first reported...reported in 1996, is an eleven β-sheet-containing “barrel” shape, with the chromophore concealed at the...molecular biology. Scientists can easily utilize GFP-containing plasmids as a means to many functional ends. ...
Showing: 451 - 500 of 809 results