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  1. Lab Automation at Addgene

    Type
    Blog Post
    ... one. As a starting point, I would recommend conferences like Lab Robotic Interest Group (LRIG). There...customizability. It has a large deck that can house many different types of devices. As long as a device fits on ... Software for Biology Tips from the Repository Trenches: Using Barcodes to Track Samples Resources at...video library Check out our molecular biology reference pages ...

  2. Tips for Titering Your Lentiviral Preps

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    Blog Post
    ...options as well as the benefits and drawbacks of different methods (for comprehensive protocols for all of...used in the transfection; if preparing several different lentiviruses in parallel, this approach should...that standard conditions be used when comparing different batches of virus. In addition, some viral preps...step of a successful transduction experiment. References 1. Kutner, Robert H., Xian-Yang Zhang, and Jakob...

  4. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

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    Blog Post
    ...without permanently altering the DNA, CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) are...study ncRNA, CRISPR systems offer a number of different options. By carefully considering your ncRNA of... you more about its function.  Resources and references Resources on addgene.org Addgene's CRISPR Guide... CRISPR enzymes for transcriptome engineering References Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna...

  5. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ...CRISPR experiment. Happy cloning! References and Resources References McGee, A. V., Liu, Y. V., Griffith...chosen because of their broad applicability in different model systems. What’s the error rate? The Doench...Table 1 has a list of Fragmid destination vectors currently available. Table 1: Fragmid destination vectors...

  6. Hot Plasmids and Viral Preps - January 2021

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    Blog Post
    ... recombinase/photodimer pairs. Created with BioRender.com In their recent ACS Synthetic Biology publication...fold more photostable and as bright to mVenus, the parent YFP, and is the most photostable YFP to date. As...and reduced RNA-off target footprint compared to current N-terminal linked SaCas9 ABE variants. It's also...specific tight affinity glutamate sensor from the Loren Looger Lab now available in AAV1. Get GFAP.SF-iGluSnFR.A184S-AAV1...

  7. 10 Basic tips for mammalian cell culture

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    Blog Post
    ...Also, keep all liquid containers closed when they aren’t in immediate use. This will prevent spills, but...could change their growth rates and morphology, rendering their continued use inadvisable for experiments...in a lab where multiple people are working on different things, this will help you avoid cross-contamination... Immunology. You can find Sana on LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling...

  8. xCas9: Engineering a CRISPR Variant with PAM Flexibility

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    Blog Post
    ...species, like S. aureus and N. meningitidis require different PAM sites than SpCas9, in this case, NNGRRT and...single PAM recognition. Created with BioRender.com. After 24 days of phage propagation, Hu ...percentages, with fold changes compared to SpCas9 in parentheses. The development of xCas9 3.6 and 3.7 shows ...is working for you in the comments section! References Hu, Johnny H., et al. "Evolved Cas9 variants ...

  9. Plasmids 101: Optimizing Plasmid Yields

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    Blog Post
    ... Let it incubate Check the bacterial strain Differences between E. coli host strains can impact plasmid...might want to include an extra wash step or use a different strain. For most applications, Addgene uses and...inadequate plasmid DNA yields. Why would volume make a difference? In the simplest of terms, more volume = more...conditions play a critical role in plasmid DNA yield. Different plasmids and strains will vary in their optimal...

  10. Anatomy of a Plasmid Page at Addgene

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    Blog Post
    ... ever found yourself bamboozled by all of the different kinds of information on our plasmid pages? Well...depositing lab, although you may have an entirely different use in mind! This section of our plasmid pages... you to use in your Materials and Methods and References sections when citing the use of your colleague...e.g. if it was derived from a plasmid made in a different lab - or one obtained from Addgene!).  It also...

  11. Plasmids 101: Degron Tags

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    Blog Post
    ...you can make an informed choice! References and Resources References Bondeson, D.P., Mullin-Bernstein,...are titratable. With these tags, protein levels aren’t limited to ‘on’ and ‘off’; varied levels of drug...profile their effectiveness with over a dozen different proteins. In the end, you can’t predict with absolute...

  13. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

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    Blog Post
    ...anymore. The kit contains sequences for several different fluorescent proteins. Visualizing fluorescence...use our promoters and terminators, but then they aren’t sure how to convert their own coding sequence to...with the Athena SWAN Charter and it has really strengthened the network of female scientists. Twitter has...for those who go to work in cancer biology.  References: A Golden Gate Modular Cloning Toolbox for Plants...

  14. Lentiviral Vector Uses and Overview

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    Blog Post
    ... applications. Read on to learn more about the current (and future) applications of lentiviral vectors...facilitates nuclear export of transcripts). A different viral envelope protein (Env), usually from VSV-G...plasmid number. For more information on the key differences between the 2nd and 3rd generation packaging ...use lentivirus in the comments section below! References 1. In vivo gene delivery and stable transduction...

  15. Isolating Genomic Regions of Interest with the CRISPR System

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    Blog Post
    ... adaptations of this system have been made in different fields of study. In addition to the identification...purification with an anti-Cas9 antibody. These different tag systems and affinity purification schemes ...dystrophy (FSHD), a disease for which there is currently no cure (Campbell et al., 2018). Find plasmids...please also see our interview with Dr. Fujii. References Campbell, Amy E., et al. "NuRD and CAF-1-mediated...

  16. Viral Vectors 101: The Retroviral Lifecycle

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    Blog Post
    ...dimerized linear ssRNA strands (which can have a different genomic sequence from one another) — by various... with lentiviral replication.  Resources and References More resources on the Addgene blog Viral Vectors...popular lentiviral plasmids γ-Retrovirus Guide References Cullen, B. R. (1992). Mechanism of action of ...Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives. Viruses, 13(7), 1288. https...

  17. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
    ...when separated (2), and that DBD and TAs from different systems could be combined effectively (3). In ...a more vigourous screen is to use a number of different bait and prey vectors. This has been shown to ...split ubiquitin system has been devised (8). References 1. Casadaban M.J., Martinez-Arias A., Shapira...Science. 1986; 231:699-704. PubMed PMID: 3080805. 3. Brent, R., and Ptashne, M. A eukaryotic transcriptional...

  18. Cpf1: A New Tool for CRISPR Genome Editing

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    Blog Post
    ...endonuclease domain, indicating that Cpf1 functions differently. Since Cpf1 loci are widely distributed across...could be adapted for genome editing. The use of a different nuclease could potentially overcome some of Cas9...Figure from Kleinstiver et al., 2016. Cpf1’s strength may lie in its low off-target editing rates, determined...shown the diversity of CRISPR-based systems. References 1. Zetsche, Bernd, et al. "Cpf1 is a single RNA-guided...

  19. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...alleles, or introduce a premature stop codon to render an RNA nonfunctional. In their quest to build a...are present in the gRNA spacer, as ADAR will preferentially edit an adenine if the template has a cytosine...(Abudayyeh et al., 2019). To avoid some of the inherent drawbacks for RNA editing of natural cytidine ... Valley contributed to updating this article. References Cox, David B.T., et al. “RNA editing with CRISPR-Cas13...

  20. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ...known quantities of DNA. Additionally, an AAV reference, a virus with a known titer, can be used to confirm...both results would be valid. This is why the AAV reference is critical. It allows you to determine if your... PCR (ddPCR) does not require a standard or a reference, meaning saving on reagents (and time)! AAV titering...becoming a popular option for PCR experiments.  References Furuta-Hanawa, Birei, Teruhide Yamaguchi, and...

  21. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    .... While these differences may seem subtle they result in a key physiological difference between lenti-... system is that it involves transfecting four different plasmids into the producer cells (two packaging...as the viral long terminal repeats (LTRs) are different. As a result, lentiviral and retroviral packaging...

  22. Negotiating Work and Life: How to Find the Joy

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    Blog Post
    ... to a conference. Really. No one will starve while you are away a few days. Also, your children will enjoy... yourself, “I’m not married,” or “I don’t have children, so my life will be under control.” Think again... hard not to do this. For a family with working parent(s), the sooner your kids can make a decent meal...

  23. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

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    Blog Post
    ...that safe-targeting guides better control for the inherent effects, or background noise, of dsDNA breaks ...on-target activity. However, there was no significant difference in depletion of ricin sensitizer genes identified...Knockout Library are available from Addgene! References 1. Morgens, David W., et al. "Genome-scale measurement...libraries Find all of the CRISPR Pooled Libraries currently available from Addgene Other Resources Watch...

  24. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...tracking experiments (Rizzo, 2009). With many different FPs that emit across the visible spectrum, researchers...of existing FP and epitope tags, researchers in Loren Looger’s lab have developed new molecular tags combining...antibodies. The “spaghetti monster” name is a reference to the deity of the social movement, the church... to the expanded use of smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance...

  25. Transferable Skills Guide: Creativity

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    Blog Post
    ...the lab can be applied anywhere! References and Resources References Grant, A. (2017). Originals how non-conformists...inclusivity. Work with and value people who think differently from you and you will arrive at unique solutions...ability to innovate. (Finkelstein, 2019) One issue currently plaguing the scientific community is supply chain...

  26. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...0-5%. This blog post covers a brief review of different basic BE technologies and their adaptation for...Nickase Cas9 (D10A) Deaminase PAM requirements Reference CBE SpCas9 rAPOBEC1 NGG 6 SpCas9 Target-AID...Rice Research Institute, Cuttack, Odisha, India. Currently Kutubuddin is a Fulbright Visiting Scholar at ...improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor, Alexis C., et al. "Programmable editing...

  27. Plasmids 101: Gateway Cloning

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    Blog Post
    ... The entry clone and destination vector carry different antibiotic resistance markers (indicated here ...STEP 1: Generate an Entry Clone There are a few different ways to generate our desired entry clone - human...interest: Find Gateway cloning plasmids here! References 1. Chee JY, Chin CF (2015) Gateway Cloning Technology... Digest Video Check Out Our Molecular Biology Reference Pages  ...

  28. Hot Plasmids: Fall 2024

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    Blog Post
    ...middle of proteins) and performs well in many different antibody-based applications.  IPI’s recombinant...virus 5 and their use in revealing antigenic differences between human, canine and simian isolates. J ...systems don’t always work across species due to differences in natural DNA repair pathways and spontaneous...at focal adhesions in living cells in several different contexts.   Figure 5: Left: Mechanism ...

  29. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

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    Blog Post
    ... the Shapiro lab at Caltech.   References and Resources References Piraner, D. I. et al. Going Deeper...produce ultrasound contrast. We (the authors) are currently working on improving and optimizing ARGs, with...straightforward to engineer various cell lines from different tissues and mammalian species to produce gas vesicles...

  30. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ...immunogenicity and range of serotypes allowing preferential infection of certain tissues. However, packaging...equally potent Cas9 protein, the Zhang lab took a different approach. They analyzed over 600 Cas9 orthologs...Sontheimer lab used NmeCas9 with AAV to edit two different disease-causing loci in mouse liver.  Two other...have also been used to express base editors. References Chew WL, Tabebordbar M, Cheng JKW, Mali P, Wu...

  31. Finding nucleic acids with SHERLOCK and DETECTR

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    Blog Post
    ...variations of Cas13 and Cas12a, are combined with different fluorescent reporters for each enzyme. In addition...Zetsche et al., 2015 ). Cas12a however recognizes a different PAM site and generates 5’ and 3’ staggered ends...Jennifer Tsang contributed to updating this article. References Abudayyeh OO, Gootenberg JS, Konermann S, Joung... Mondini A, Nogueira ML, Isern S, Michael SF, Lorenzana I, Yozwiak NL, MacInnis BL, Bosch I, Gehrke L,...

  32. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    Blog Post
    ... Optimizing FLP FRT through smart engineering Different FLP versions – making FLP efficient The first ...efficiency to Cre (Kranz et al. 2010). These differences in recombinase efficiency are important to keep...should be visualized at the same time from two different plasmids this approach becomes problematic as ...the genOway press release for more information. References 1. Atasoy, D., Y. Aponte, H. H. Su, and S. M....

  33. 5 Reasons to Use Reddit for Science Communication

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    Blog Post
    ...of conversations with people who work in many different scientific fields. They also host AMA forums where...first makes Reddit a great platform for those who aren’t so great at coming up with answers off the cuff...future. The AMA was also cataloged for permanent reference by the Winnower. The only small downside to the...leaving out the term “implicit bias” which is apparently a troll trigger phrase these days). Redditors...

  34. Filming Science Videos in the Age of Social Distancing

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    Blog Post
    ... involved multiple people in tight locations. Conference rooms for on-screen presenters! Labs and benches...Zoom, everyone’s new favorite third-party video conferencing software. Interviewees could simply boot up ...while the camera is here, the operator is in a different room.   Design: The previous cycle made me...monitor and teleprompter being repurposed for a different shoot to create the How to Deposit Plasmids video...

  35. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ...testing is crucial in controlling this pandemic. Currently, the CDC uses RT-qPCR tests to diagnose COVID-...Cas12a, Cas13a does not require strict sequence preferences at the target site, while Cas12a requires a PAM...femto-aptomolar range. For more details on the differences between these methods, check out this table.  ...can follow her on Twitter @shra_krishna.       References Azhar, M., Phutela, R., Ansari, A. H., Sinha,...

  36. Technologies Enabled by NanoLuc® Luciferase

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    Blog Post
    ...more common luciferases like firefly (FLuc) and Renilla (RLuc) luciferase. This is important not so much...strategy was first applied using RLuc8, an engineered Renilla luciferase, to produce a series of FP-RLuc8 fusions... #85201), Orange (OeNL; plasmid #85202) and Red (ReNL; plasmid #85203) NLuc fusions. The Nagai lab has...Promega cellular analysis products at Promega.     References 1. Hall, M.P., et al. (2012) Engineered luciferase...

  37. Technique: Probe Phage Genomes for Host Binding Proteins

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    Blog Post
    ...influence the evolution of their bacterial prey. Currently, receptor binding proteins (RBPs, the host recognition...thinking. Find her on Twitter @JessicaSacher.   References 1. Nobrega, Franklin L., et al. "Revisiting phage...phage therapy: new applications for old resources." Trends in microbiology 23.4 (2015): 185-191. PubMed PMID...bacteriophage-encoded glycan binding proteins and carbohydrates." Current opinion in structural biology 34 (2015): 69-77....

  38. CRISPR 101: Epigenetics and Editing the Epigenome

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    Blog Post
    ...with transcriptional activation. Created with BioRender.com. Tet1 demethylase Ronggui Hu’s lab has created... with increased transcription. Created with BioRender.com. Tet1 initiates cytosine demethylation of...with transcriptional repression. Created with BioRender.com. DNA Methyltransferase MQ1 Margaret Goodell's...Schwiesow contributed to updating this article. References Haldeman JM, Conway AE, Arlotto ME, et al (2018...

  39. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
    ... strategies in C. elegans As brilliantly and irreverently illustrated in the parable of the Geneticist...the sequence added, leaving a 34 bp “scar”, but currently requires additional experimental manipulation....days. The only animals on a plate, other than the parental animal, are rescued progeny, which makes screening...more at his webpage or follow him on Twitter.   References Arribere JA, Bell RT, Fu BXH, Artiles KL, Hartman...

  40. Which Fluorescence Microscopy Technique is Best for Me?

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    Blog Post
    ...excitation light path. Each hole corresponds to a different location in the sample. As more than one hole ...one of four categories that are each suited to different types of modern day fluorescence microscopes (...means that the images will appear sharp without interference from blurry out-of-focus light. The most difficult...microscopy manufacturers have similar information References 1. Lichtman JW, Conchello JA. Fluorescence microscopy...

  41. Find and Share AAV Data with Addgene's New AAV Data Hub

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    Blog Post
    ...over 100 entries spanning six different species and dozens of different expression sites. The Data...the AAV Data Hub is a beta project, and we are currently only accepting data for materials distributed ...

  42. Call for Guest Bloggers

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    Blog Post
    ...has deposited If you’d like to write about a different topic, feel free let us know. What do you get ...audience of biological scientists from a variety of different fields. Exposure - With over 100,000 views each...interact with and discuss your work with many different people. Recent Guest Writers Have Included: Wenning...

  43. Antibody Data Repositories and Search Engines

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    Blog Post
    ...search engines, which allow you to search many different antibody catalogs at the same time.  If you're...Library for Cancer Research Human Luekocyte Differentiation Antigen Workshops Immune cells Flow cytometry...others) Data repository From the Human Cell Differentiation Molecules group Addgene Data Hub Any ...

  45. Interview with Bart Newland, Addgene’s Newest Board Member

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    Blog Post
    ...demand. Q: Can you tell me a bit about your job currently? What is your day-to-day like? BN: I've made the...large companies, Genzyme and Biogen. They’re very different jobs. Being general counsel at a small start-up...companies. It's one of those "every day is something different" type of jobs, which really suits my personality...

  46. Kwabena Duedu on Public Health, Biofuels, and Doing Science in Ghana

    Type
    Blog Post
    ... useful products under Professor Christopher E. French. Dr. Duedu was recently awarded plasmids from Addgene...7:12 - 7:11 : Kwabena's work in Christopher E. French's Lab at the University of Edinburgh - synthetic...bioresource to attract research funding. Using current resources to develop microbiology and molecular...

  48. Fueled by Coffee at #SfN14

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    Blog Post
    ...more than 30,000 neuroscientists over a 5 day conference? I feel like here at the annual Society for Neuroscience...late, late night socials. As a newbie to the SfN conference, I have been impressed by how the neuroscience...reports from bloggers at the meeting – my report is currently in the works. Or follow #SfN14 on Twitter for ...

  49. What's New in CRISPR - May 2019

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    Blog Post
    ...AAV constructs CRISPR interference in Candida albicans A new CRISPR interference (CRIPSRi) platform for... to Cas9 SECURE base editors Nme2Cas9 CRISPR interference in Candida albicans E. coli genome-wide inhibition...

  50. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    Blog Post
    ...validated in.  If a plasmid's Addgene ID # has been referenced in other publications, you'll find a link to ...Additional features Once you've clicked on the "# References" link under the "Resource Information" heading...Addgene’s Plasmids 101 eBook Protocols and Plasmid References @Addgene  ...
Showing: 651 - 700 of 1014 results