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Showing: 281 - 320 of 621 results

  1. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...designing primers, and running analyses. New team members have taken up to a month to design a single 96-...began building CrispyCrunch in July 2018 and in November, we began using it internally. Guiding principles...biopython, bwa, samtools, bowtie, UCSC genome browser, Ensembl, and others. It builds on existing popular web ...

  2. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...it may benefit you to work as a team with other members of your lab, a lot of the “soft skills” required... In an academic environment, surrounded by lab members who are all working on related topics, scientific...will be part of a team in industry with most team members giving oral presentations, you will also have to...

  3. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ... (nr/nt)” as it contains a composite of GenBank, EMBL, DDBJ, and PDB sequences and may be the most comprehensive...regions by default, meaning that you do not have to remember to adjust any blastp algorithm parameters. We ...http://www.expasy.org/genomics/sequence_alignment EMBL-EBI - http://www.ebi.ac.uk/services http://www....

  4. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...figure above), which generates pores in the cell membrane, allowing for entry of the RNP into the cytoplasm...nucleofection, which forms pores in the nuclear membrane, allowing for entry of a DNA template. Other techniques...protoplasts and biolistic bombardment of immature embryos (Liang et al. 2017).  PEG-mediated methods, while...

  6. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    ...notebooks and computer files. If you asked your team members or colleagues: How much unpublished AAV data do...Therapy - Methods and Clinical Development, 18(September), 98–118. https://doi.org/10.1016/j.omtm.2020.05.018...Stimulation. Frontiers in Human Neuroscience, 14(December), 1–12. https://doi.org/10.3389/fnhum.2020.578687...

  7. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...compromising the photon budget. Likewise, the switch from ensemble imaging to single-molecule imaging places a greater...recently discovered that incorporation of four-membered azetidine rings could substantially enhance the...analogs without sacrificing their small size and membrane permeability (Figure 2c). These cell-permeable...

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

    Type
    Blog Post
    ...data from the ENCODE project, find enhancers in embryonic stem cells, and conclude with a targeted CRISPR...for NANOG, an essential transcription factor in embryonic stem cells (ESCs). Your ChIP-seq finds just over...CRISPR screen. We start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an...

  9. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...development, complex genetic programs specify and assemble diverse arrays of neurons, forming the neuronal...subgenome-scale CRISPRa screening approach in embryonic stem (ES) cells to determine which transcription...gene-function relationships in the CNS using CRISPRa as a member of the Gelbard Laboratory at URMC.     References...

  10. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ... The G·U wobble base pair. EMBO Rep 1:18–23 . https://doi.org/10.1093/embo-reports/kvd001 Additional resources...mammalian cells. This was the first codon optimized membrane protein expressed in mammalian cells. In this ...

  11. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    ...for attendees and helping societies help their members get the most out of their meetings. Amanda Gunn...Gunn ran randomized coffee trials for us at ASCB|EMBO 2019, matching Mentors with Singletons for short coffee... a culture that brings more value to ALL their members.  Shoutout to our mentors and supporters Thanks...

  12. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...structures in RNA transcripts Effects on transcription (remember, it’s not all about translation - e.g. codon choice... isoacceptors induced by amino‐acid starvation." EMBO reports 6.2 (2005): 151-157. PubMed PMID: 15678157...transfer RNA abundance in Escherichia coli." The EMBO journal 9.13 (1990): 4359-4366. PubMed PMID: 2265611...

  13. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ... streptavidin-coated magnetic beads through an assembly of biotinylated SpyTag-SpyCatcher linkers and ...and target capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes...loss. Ci Ji Lim’s lab found that adding Late Embryogenesis Abundant (LEA) proteins, small proteins that...

  14. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ...heads and long legs that protrude from the cell membrane, physically connecting the extracellular environment...the integrin, where it is resisted by the ligand embedded in the extracellular environment. This further...the extended open integrin conformation. During embryonic development, extended-open integrins enable tissue...

  15. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA...they observe fluorescence localizes to the plasma membrane only when the RTK is activated and the ITAM phosphorylated...relies on a change in localization to the cell membrane and not a direct change in the fluorescent properties...

  16. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...hydroxybenzylidene imidazolinone (DFHBI). DFHBI is membrane permeable and non toxic to cells. Because it rapidly...split aptamer that can be used to investigate RNA assembly. The split aptamer is composed of a “Top” and ...fluorescent split aptamer for visualizing RNA–RNA assembly in vivo." ACS synthetic biology 6.9 (2017): 1710...

  18. Trimmer Lab NeuroMab Collection

    Type
    Collection
    ...translocation of the heavy chain across the ER membrane. PCR-mediated fusion of these three elements is...translocation of the light chain across the ER membrane. Downstream of the insert is a heavy chain constant...

  19. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...deposited versions with both PDGFR and NGR fusions for membrane display, they recommend the NGR vectors for most... modulators of cell signaling pathways during embryonic development and cancer. But there is still much...intestinal peptide (VIP), or lysosomal-associated membrane protein 5 (LAMP5) — as well as cholinergic (choline...

  20. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...optogenetic accuators to depolarize or hyperpolarize membranes. Traditional FRET reporters could not be used ...Venus fluorescent protein in conjunction with a membrane-inserted voltage sensing domain (LOTUS V; Luminescent...Universal Sensing of Voltage). With this sensor, membrane depolarization increases the BRET signal whereas...

  21. A Guide to Getting Started in Undergrad Research

    Type
    Blog Post
    ... scientific mentor and choosing the right lab. Remember that your time is valuable! Don’t jump on the ...to plan out a very distinct and certain path, embracing uncertainty can help you both in your career and....org/10.1126/science.aba8039 Cordova FA (2016) Embrace uncertainty. Science 351:994–994 . https://doi....

  22. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...committee in graduate school. Thesis committee members provide expertise in designing and interpreting...to introduce yourself to and interact with the members of your field, and present your research. Face ...family. Those people are your support network, so remember to thank them verbally and often for their love...

  23. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...SDS-PAGE, and are subsequently transferred to a membrane. A primary antibody recognizing the protein of...is added, and will bind to that protein on the membrane. Then, addition of a secondary antibody allows...bind to proteins on the cell surface, or the cell membrane can be permeabilized to allow antibodies to bind...

  24. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    ...speed at which a confocal image can be acquired. Assembling an image pixel by pixel is slow; therefore, certain...detector such as a scientific CMOS camera. If plasma membrane dynamics are the main focus of the experiment,...dynamic samples Ex: 3D cell cultures, small model embryos >15 μm Thsee are some of the most challenging samples...

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

    Type
    Blog Post
    ... in CRISPR–Cas9. The EMBO Journal, 37(10), e96941. https://doi.org/10.15252/embj.201796941 Pattanayak,...CTD and the entire REC lobe—do not structurally resemble any other known proteins; they are completely ...

  26. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...single stranded, negative sense RNA virus. It’s a member of the Paramyxoviridae family of viruses, which...staining for alkaline phosphatase, expression of embryonic proteins, and promoter demethylation and expression.... Cells can be differentiated in vitro to form embryoid bodies which are compact balls of loosely organized...

  27. Deep Dive: qPCR

    Type
    Blog Post
    ...1), which may be easier to conceptualize. Just remember to switch them back before reporting the data!...use it! Tempting as it may be to skip this step, remember we here at Addgene frown upon statistical laxness...11237753. Primer design resources PrimerBlastDINAmelMfoldBLASTEnsembl More resources on the Addgene blog...

  28. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ... Biology 22:169–174 . https://doi.org/10.1016/j.chembiol.2014.12.011 Polstein LR, Gersbach CA (2015) A...Chem Biol 11:198–200 . https://doi.org/10.1038/nchembio.1753 Richter F, Fonfara I, Bouazza B, Schumacher...Chem Biol 13:443–448 . https://doi.org/10.1021/acschembio.7b00603 Additional Resources “Optogenetics Meets...

  30. Lucky Thirteen: Guest Contributors of 2023

    Type
    Blog Post
    ...It's December, and that means it's officially the start of Addgene's winter holiday celebrations! We'...with Addgene writers High Complexity Golden Gate Assembly with NEB's DADExperts: Matthew Norton and NEBGCE4All...

  31. SciComm with the Experts at Science in the News Part 1

    Type
    Blog Post
    ... former Co-director of Science in the News, and member of the SITN podcast team.     Amy Gilson recently...former Co-director of Science in the News, and is a member of the SITN podcast team. You can follow Amy on...

  32. Depositor Week 2020 is Happening Now. Join us!

    Type
    Blog Post
    ...Mark your calendars for November 16-20, 2020 for #DepositorWeek on Twitter!  That’s today through Friday...presented in this blog post includes data until November 9, 2020. Between then and the posting of the blog...

  33. Celebrating One Million Plasmids Shared!

    Type
    Blog Post
    ... that everything will ship out in time - from assembling the boxes, to inoculating tubes, and to packing...pic.twitter.com/eJbI0lLiGQ — Dr. Joanne Kamens (@JKamens) September 13, 2018 This massive scale reagent sharing led...

  36. MassBioEd Career Exploration Day at Addgene

    Type
    Blog Post
    ...intellectual property from our Tech Transfer team members Tony Cova and Chiara DeNatale and programming with...great European photo slideshow from our UK team member Benoit Giquel. Bluegene’s globetrotting impressed...

  38. Keeping Up With CRISPR/Cas9

    Type
    Blog Post
    ...are coming along at a fast and furious pace. In December, as we've blogged about before, there was the ...via Cas9/RNA-Mediated Gene Targeting in One-Cell Embryos. Niu Y, et al. Cell. 2014 January 30. (Pubmed) ...
Showing: 281 - 320 of 621 results