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  1. How to Be an Excellent Trainee

    Type
    Blog Post
    ...advice, open doors for you, and teach you valuable lessons along the way. We think it’s so important that ... technician, or a graduate student. This is the person you report or talk to on a daily basis who will...scared to ask for some! You can email or ask in-person “Are there any papers you think would be useful...these things will help you get more out of the in-person training component. Documenting during training...take on trainees one day when you are the senior personnel! Happy training! Resources Resources on the Addgene...
  2. CRISPR Activation: A Practical Guide

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    Blog Post
    ...Pinnelli M, Ansari R, Harper S, Jackson DA, McRae R, Pooley R, Wilkinson P, van der Meer D, Dow D, Buser-Doepner...of TSS for best results but up to 400 bp works reasonably well.  Best of luck on your experiments!  Many...Housden BE, Perrimon N, Collins JJ, Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods...doi.org/10.1038/nmeth.3871 Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes...
  3. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...favorably to other methods. In a head-to-head comparison with Cas9, Cas9 modified 9/12 known off-targets... C., Rees, H. A., Packer, M. S., Badran, A. H., Bryson, D. I., & Liu, D. R. (2017). Programmable base ...doi.org/10.1038/nbt.3803 Koblan, L. W., Doman, J. L., Wilson, C., Levy, J. M., Tay, T., Newby, G. A., Maianti... Bohnuud, T., Fernandez, T. P., Gehrke, J. M., Grayson, P., Lee, K. D., Ortega, M. A., Sawyer, C., Schwaegerle..., Lapinaite, A., Newby, G. A., Thuronyi, B. W., Wilson, C., Koblan, L. W., Zeng, J., Bauer, D. E., Doudna...Levy, J. M., Yeh, W.-H., Zheng, C., Newby, G. A., Wilson, C., Bhaumik, M., Shubina-Oleinik, O., Holt, J....
  4. Five Popular Model Organisms

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    Blog Post
    ...cells are great model organism not only for the reasons above but because they actually share many biological... the study of neurological diseases, such as Parkinson’s disease (PD). PD is primarily characterized by...pancreas and produce different levels of insulin (Hesselson et al., 2009). The worm (Caenorhabditis elegans...nature391.6669 (1998): 806. PubMed PMID: 9486653. Hesselson, Daniel, et al. "Distinct populations of quiescent...From the baker to the bedside: yeast models of Parkinson's disease." Microbial cell 2.8 (2015): 262. PubMed...
  5. Antibodies 101: Single Chain Fragment Variables (scFvs)

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    Blog Post
    ...10.3390/antib8020028  Bird R, Hardman K, Jacobson J, Johnson S, Kaufman B, Lee S, Lee T, Pope S, Riordan... (Sandomenico et al., 2020). Figure 1: Comparison between the IgG antibody and scFv.   What...doi.org/10.1371/journal.pone.0189964  Huston JS, Levinson D, Mudgett-Hunter M, Tai MS, Novotny J, Margolies... made by molecular evolution 1 1Edited by I. A. Wilson. Journal of Molecular Biology 275:245–253 . https...
  6. Viral Vectors 101: AAV Variables That Matter

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    Blog Post
    ...designing and testing your AAV.  Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8, and 9 in two ...Taschenberger et al., 2017).     Figure 3: Comparison of two astrocyte selective promoters demonstrating...Sortwell, C. E., Hauswirth, W. W., Boye, S. E., & Manfredsson, F. P. (2017). Rationally Engineered AAV Capsids...vector-mediated transduction of the corticospinal tract: Comparison of four promoters. Gene Therapy, 28(1–2), 56–... Jason Nasse...
  7. Plasmids 101: Dimers and Multimers

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    Blog Post
    ...PMC2693335. Williams JA, Luke J, Langtry S, Anderson S, Hodgson CP, Carnes AE. Generic plasmid DNA production...and at high copy numbers (Williams, Carnes, and Hodgson, 2009). More research is ongoing to determine plasmid...90057-7. PMID: 2544913. Cook KS, Luo J, Guttman A, Thompson L. Vaccine Plasmid Topology Monitoring by Capillary...90060-6. PMID: 6323019. Williams JA, Carnes AE, Hodgson CP. Plasmid DNA vaccine vector design: impact on...
  8. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
    ...2013), or rescue of mutant phenotypes (unc-119) (Dickinson et al., 2013). These approaches allowed effective...the development of the self-excising cassette (Dickinson et al., 2015) made this step as simple as heat-shocking...e193–e193 . https://doi.org/10.1093/nar/gkt805 Dickinson DJ, Pani AM, Heppert JK, Higgins CD, Goldstein...sequences https://doi.org/10.17912/W2237D Farboud B, Severson AF, Meyer BJ (2018) Strategies for Efficient Genome...Krishnamurthy PM, Gneid H, Edraki A, Watts JK, Sontheimer EJ, Mello CC (2018) 5′ Modifications Improve ...
  9. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    Blog Post
    ...many stages of synthesis and degradation. For this reason it is virtually impossible to determine specific...parameter to consider is the contrast ratio- a comparison of PA-FP brightness before and after photoactivation...extraordinary molecular detail.   References 1.Patterson, George H., and Jennifer Lippincott-Schwartz. ..., Samuel T., Thanu PK Girirajan, and Michael D. Mason. "Ultra-high resolution imaging by fluorescence ...Photoactivatable FPs in Empty Backbones in the Michael Davidson Collection Find Photoactivatable FPs for Single...
  10. Editor's Choice, September 2016

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    Blog Post
    ...contributed by Kurt Swanson, husband of Addgene Director of Biology, Lianna Swanson, and researcher at ...in food products). I picked this post for a few reasons: 1) I think phage are super cool and have considered...proteins that could be useful for a wide variety of reasons. 3) I would love to see more posts like this on...
  11. Bricking Science: Portraying Scientific Reality Through LEGO

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    Blog Post
    ... spend a few days editing and later looking for songs and sounds to synchronize with the images thereby...the hardest part of the process was to embed personality in pieces of plastic, but it’s gotten easier ...than a piece of plastic for us now). With the personality in place, everything now is about the plot. This...How You Can Use Video to Share Your Research 5 Reasons to Use Reddit for Science Communication 10 Ways...
  12. Management for Scientists: Giving Feedback

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    Blog Post
    ...to improve, and they’re also clear about how the person is doing overall. Employees hate to wonder what...employee. Make sure to focus on the action, not the person - intent is usually good. Don’t pretend something...standard format not only enables you to discuss a person's performance or what has been learned, it helps... effective over time if you take it seriously.  Person who will receive feedback does a self-assessment...
  13. Choosing a Good Mentor for Scientists

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    Blog Post
    ... ask a lot of questions. Personality and style conflicts are a common reason for scientists to change ...budgeting, equipment acquisition, public speaking and personnel management?  If you are thinking about a non-academic... you can help it. Observe as much as you can in person. Rotate in the lab, if possible. Attend a lab meeting...
  14. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...use Bioluminescence Resonance Energy Transfer (BRET) were developed by the Johnson Lab . Characteristics...CRISPR labeling chromosomal loci in genome Thoru Pederson’s lab at the University of Massachusetts Medical...lab further modified sdChr- a blue-shifted channelrhodpson from freshwater green alga identified by Ed...Nano-lanterns rely on the principle of bioluminescence resonance energy transfer (BRET), whereby photons emitted...-tagged human ORF clones to allow testing and comparison of different cell-free expression systems; the...understanding complex gene regulation (Bert et al 2000, Johnson et al 2004). So if you are looking for a luciferase... et al. Plasmid. 2000. PubMed PMID: 10964627 Johnson, et al. Mol Cell Biol. 2004. PubMed PMID: 15340054...
  15. What Do I Do Now? Academic v. Non-Academic Career Decisions

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    Blog Post
    ...   In fact, there are a lot of good reasons to stay and good reasons to go.  Here are some questions and...especially race and gender) Interpersonal issues can create interpersonal issues (one bad apple can change...
  16. Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring

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    Blog Post
    ...process samples at the picnic table before dark, a bison wandered into our campsite. We had to abandon our...our samples and move farther away until the bison meandered off. In the end we were able to safely get...get back to our lab work. Figure 3: When a bison wanders into the campsite, leave your samples behind...ThermoCas9 from the van der Oost lab. Phylogenetic comparison indicates that like GeoCas9 and ThermoCas9, IgnaviCas9...
  17. The Strength of Story Telling

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    Blog Post
    ...end; cause, effect; lesson, learning. Storytelling is a crucial part of interpersonal communication, which... it came time to add author details a different person in the lab wanted to be the first author. When ...protagonist will be you. Make sure to highlight what lessons you’ve learned and what knowledge you’ve gained...
  18. Another Pathway into Cells: iTOP

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    Blog Post
    ...Although CRISPR/Cas9 is all the rage, they had another reason to test this system. It’s difficult to determine...require multiple transduction events. For these reasons, iTOP is best suited to binary systems in which... size associated with viral vectors. For these reasons, iTOP, especially in tandem with genome editing... VP22 nuclear homing an artifact? Lundberg M, Johansson M. Nat Biotechnol. 2001 Aug;19(8):713-4. PubMed...
  19. Antibody Validation for Flow Cytometry

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    Blog Post
    ... T. M., Edwards, A. M., Healy, L. M., Robertson, J., & McPherson, P. S. (2019). Implementation of an antibody...perform as well in other conditions. For these reasons, it is essential to test the selectivity of your...cannot prove antibody selectivity, and for that reason, it is best practice to combine correlation with...C., Bandrowski, A., Virk, H., Edwards, A. M., McPherson, P., & Laflamme, C. (2023). Scaling of an antibody...
  20. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

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    Blog Post
    ...Thanks to our guest blogger Cameron MacPherson! Cameron R. MacPherson is Lead Data Scientist for the Milieu...post was contributed by guest blogger Cameron MacPherson at the Institut Pasteur CRISPR software and the... by their focus on specific organisms. For this reason I generally recommend that laboratories that have...CRISPR technology to a new organism: For obvious reasons you can rule out any tool that limits you to a ...
  21. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

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    Blog Post
    ...fluorescent proteins everyday.  The late Michael Davidson and Roger Tsien played enormous roles in enabling...their legacy, we recently launched the Michael Davidson and Roger Tsien Commemorative Travel Awards. These... we’re proud to announce our first two Michael Davidson and Roger Tsien Commemorative Travel Awardees:...Plasmids from the Tsien Lab Browse the Michael Davidson Collection ...
  22. Addgene Depositors Get More Citations

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    Blog Post
    ...contributed by guest bloggers Samantha Zyontz and Neil Thompson from the MIT Sloan School of Management. Professor...depositing a plasmid with Addgene, in contrast to other reasons for the correlation. However, determining this ...to our guest bloggers, Samantha Zyontz and Neil Thompson. Special thanks to Addgenies Pamela McA'Nulty ...page: https://ssrn.com/author=1232466.     Neil Thompson is an Assistant Professor of Innovation and Strategy...
  23. Using CRISPR/Cas9 to Edit Disease Out of the Genome

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    Blog Post
    ... by Kendall Morgan and updated in 2022 by Lucie Wilson. Lucie is an Addgene co-op from Northeastern University...in monoallelic diseases In 2013, a team led by Jinsong Li from the Chinese Academy of Sciences found that...fibrosis (8).   The difference with CRISPR/Cas9 in comparison to other genome editing techniques is its ease...s Disease.” Journal of Alzheimer's disease & Parkinsonism. https://doi.org/10.4172/2161-0460.1000439 6...
  24. K. phaffii: Rising to the Occasion

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    Blog Post
    ...make for an excellent model organism for a few reasons: Affordable to culture and easy to work with ...established in yeast, including a widely used Parkinson’s experimental model. In fact, approximately 25%...of the uses outlined below. Biology They say comparison is the thief of joy, but I think it’s acceptable...aren’t going anywhere anytime soon, and for good reason! However, there are other players in the game (...
  25. Four Factors that Differentiate the Stem Cell Field

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    Blog Post
    ...When our Director of Biology, Lianna Swanson, suggested that I might be able to attend the 10 Years of...degeneration; Laurenz Studer discussed his work with Parkinson's Disease; Hideyuki Okano presented ground-breaking...clinical trials for ALS.   I have an uncle with Parkinson's, a half-brother who is a quadriplegic due to ... are key to the research, but they are also the reason this research is happening. Stem cells scientists...
  26. Synthetic Photobiology: Optogenetics for E. coli

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    Blog Post
    ...Levskaya A, Chevalier AA, Tabor JJ, Simpson ZB, Lavery LA, Levy M, Davidson EA, Scouras A, Ellington AD, Marcotte...Chemical effectors are suboptimal for a number of reasons, including their potential toxicity and cross-reactivity...genetic edge detection program. Tabor JJ, Salis HM, Simpson ZB, Chevalier AA, Levskaya A, Marcotte EM, Voigt...
  27. Management for Scientists: Delegating is Key

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    Blog Post
    ...come into being through the actions of a single person but through the coordinated efforts of many individuals...collection page, we certainly don't expect that one person will write all the copy, design every icon, curate...page. That would be entirely too much work for one person, it would take forever, and the end product would...who can do the job. It is good to delegate so the person doing the task will have to learn and stretch a...
  28. Sweating the Small Stuff: Details in the Lab

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    Blog Post
    ...in the form of a protocol, and you can add in personalized steps like “check drawers for light-sensitive....      References and Resources Images used Roberson Lab, UPenn. Western blotting. Retrieved Feb. 7... 7, 2022. https://www.med.upenn.edu/robertsonlab/assets/user-content/documents/Western%20Blotting.pdf  ... Rachel Leeson...
  29. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

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    Blog Post
    ...PMID: 29553576. 2. Kittleson, Joshua T., Sherine Cheung, and JChristopher Anderson. "Rapid optimization..., and ColE2 plasmids) as cell division slows (Kittleson et al., 2011). Segall-Shapiro et al. showed that...References 1. Segall-Shapiro, Thomas H., Eduardo D. Sontag, and Christopher A. Voigt. "Engineered promoters...
  30. Fluorescent Proteins 101: Introduction to FRET

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    Blog Post
    ...    What is this FRET you speak of? Förster Resonance Energy Transfer (FRET) was originally described...has led to the alternative acronym Fluorescence Resonance Energy Transfer. Unlike the typical excitation...Müller, Sara M., et al. "Quantification of Förster resonance energy transfer by monitoring sensitized emission... Jason Niehaus...
  31. A Guide to Designing a Scientific Poster: Content, Layout, and More

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    Blog Post
    ...also probably thought about why certain posters resonate with you (or not). It is important to reflect ...length, and will be more difficult to read. The Thompson Rivers University Library provides useful tips...scientific poster design was conceived by Mike Morrison, a psychology doctoral student at Michigan State... avoiding red and green is a good idea for this reason but there are many other ways to make your poster...
  32. Making CRISPR Plasmids Using Fragmid

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    ...AAV pRDA_889 (ITRs from AAV2) Mammalian Transposon pRDA_757 (Piggybac; constitutive expression... drosophila) Delivery type (lentivirus, AAV, transposon, Plasmid) Destination vector (dropdown options..., Bosch, J. A., Zirin, J. D., Viswanatha, R., Sontheimer, E. J., Goodale, A., Greene, M. A., Green, T.... Rachel Leeson...
  33. Gaining Leadership Skills Volunteering at a Professional Organization

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    Blog Post
    ...national levels. These skills will also help the person mentor others more successfully. But how does one...phone calls. This has its challenges as meeting in person is often much more productive and makes it easier... previous formats but I learnt to overcome the personal frustration and submit reports in the format requested...voice may not always be perfectly aligned with my personal viewpoint but, as a leader in the NPA, I had to...
  34. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

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    ...proteins on the cell surface. Figure 1: Comparison of common antibody, HCab, and nanobody. For more...starting library size (up to 1012 members) in comparison to other display systems using phage and yeast...Egloff P, Hutter CAJ, Kuhn BT, Bräuer P, Newstead S, Dawson RJP, Geertsma ER, Seeger MA (2020) Generation of... Gera J, Gmür S, Spies P, Gygax D, Geertsma ER, Dawson RJ, Seeger MA (2018) Synthetic single domain antibodies...
  35. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

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    ...edit, it simply becomes impossible (or at least unreasonable) to test them all. This is where PRIDICT comes...find their way into the hands of users. For this reason, we made our tools available via the PRIDICT website...A. A., Koblan, L. W., Levy, J. M., Chen, P. J., Wilson, C., Newby, G. A., Raguram, A., & Liu, D. R. (2019.... M., Pallaseni, A., Kuzmin, I., Raudvere, U., Peterson, H., Liberante, F. G., & Parts, L. (2023). Prediction...
  36. Capturing a Moment with Marc Zimmer’s “The State of Science”

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    ...the most successful scientists have outgoing personalities to supplement their intellectual prowess and... era of social media and self-promotion, and personality is playing an increasingly important role in ... that produced by the confident and flamboyant Watsons.” In Part Four, “New Science,” Zimmer tackles more...little bit about many different things (part of the reason Addgene has been such a great fit for me!), the...
  37. Antibodies 101: Producing Recombinant Antibodies

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    ...PMID: 31891733. Brechmann NA, Schwarz H, Eriksson PO, Eriksson K, Shokri A, Chotteau V. Antibody capture....106. PMID: 33151027; PMCID: PMC7665837. L'Abbé D, Bisson L, Gervais C, Grazzini E, Durocher Y. Transient...Chen E, Haidar JN, Srivastava A, Goldstein J. Comparison of predicted extinction coefficients of monoclonal...
  38. Viral Vectors 101: The Retroviral Lifecycle

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    ... yet retroviruses are RNA-containing viruses  (Robinson et al., 1965)! You may be beginning to realize...Brian O'Neill, with thanks to Addgenie Rebecca Hutcheson for her assistance with lentiviral replication...PMID: 26225354. PubMed Central PMCID: PMC4515362 Robinson, W. S., Pitkanen, A., & Rubin, H. (1965). The ... PMCID: PMC355630 Faras, A. J., Taylor, J. M., Levinson, W. E., Goodman, H. M., & Bishop, J. M. (1973)...
  39. Negotiating Work and Life: How to Find the Joy

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    ...maximise the time you are enjoying both work and personal life. There are no easy answers to negotiating...my colleagues and me. To that end, here are my personal 10 commandments for making it all work – most ...the “jobs” that take you away from having your personal time. Someone else can clean your house, pick ...you can’t make it to dinner this week. One swim lesson missed can mean a world of mental relaxation. And...
  40. Fluorescent Proteins 101: Aptamer Fluorophores

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    ...fluorescent biosensors. Aptabeacons Fluorescence resonance energy transfer (FRET) technology has been studied... sensitive detection assay. In 1999, Grate and Wilson paired laser cleavage and an RNA aptamer that bound... May 15;26(9):3725-36. PubMed PMID: 21419619. 9. Song, Kyung-Mi, et al. “Aptamers and Their Biological... 22368488. PubMed Central PMCID: PMC3279232. 10. Song, Shiping, et al. “Aptamer-based biosensors.” TrAC...
  41. Save Time with Transient Plant Leaf Transformations

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    ...biologist some time… months, in fact. In the Lee-Parsons lab, we are investigating how the production of...protein of interest. Find all plasmids from the Lee-Parsons lab. Test different leaf infiltration methods ...(Germany), before starting his work in the Lee-Parsons Lab.     References D'Aoust, Marc-André, et al.... De Ruijter, N. C. A., et al. "Evaluation and comparison of the GUS, LUC and GFP reporter system for gene...
  42. Viral Vectors 101: Pseudotyping

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    ...host tropism of your virus, there are many other reasons when you might pseudotype your virus:  Lowering...a cell surface to endocytose into the cell. The reason VSV-G is so commonly used is that it is good at...Sandoval, 2017). This high titer is one of the reasons VSV-G is used to transduce stem cells and neurons...https://doi.org/10.1093/hmg/10.19.2109 Sandrin V, Boson B, Salmon P, Gay W, Nègre D, Le Grand R, Trono ...
  43. Overcoming the AAV Size Limitation for CRISPR Delivery

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    ...10.1038/nature09523  Ibraheim R, Song C-Q, Mir A, Amrani N, Xue W, Sontheimer EJ (2018) All-in-one adeno-associated...can be turned off by anti-CRISPR proteins. The Sontheimer lab used NmeCas9 with AAV to edit two different...0  Kim E, Koo T, Park SW, Kim D, Kim K, Cho H-Y, Song DW, Lee KJ, Jung MH, Kim S, Kim JH, Kim JH, Kim ...
  44. Transferable Skills Guide: Creativity

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    ...relevant to you. Elect a devil’s advocate. This is the person whose job it is to disagree with the group. They...sure we have all experienced a meeting where one person speaks up after a group consensus and is immediately...entire company. It is important to note that the person who submitted the idea did not have to be in charge...always done it this other way,” you probably have reason to continue advocating for your idea. Our product...
  45. Plasmids 101: Stringent Regulation of Replication

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    ...restraining further replication.   Figure 3: Comparison of replication regulation mechanisms of stringently...regulation! This post was originally written by Jason Niehaus in December 2015 and was updated by Emily...464.1998 Grimwade, J. E., Rozgaja, T. A., Gupta, R., Dyson, K., Rao, P., & Leonard, A. C. (2018). Origin recognition.../doi.org/10.1093/nar/gky457 Park, K., Han, E., Paulsson, J., & Chattoraj, D. K. (2001). Origin pairing...
  46. March for Science

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    ...believe GMOs are safe for human consumption in comparison to 37% of the general public (5, 6). Similarly... the public (8–13). Facing the possibility that reason is losing ground in public discourse, many scientists...politicians to interpret it for them. Other popular reasons to march include: General inclusion of science...policy should have more weight than how any one person feels about it. And yet research shows that thoughts...
  47. Tips for Getting a Faculty Position

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    Blog Post
    ... make the best case for your candidacy. A major reason that a lot of candidates either get no interviews...understand your cover letter and research statement. My personal litmus test was whether my wife could explain ... please introduce yourself to me by email or in person. If you feel compelled to repay me in some way,...job searches. If you feel the urge to repay me personally, I will gratefully accept a very hoppy pint of...
  48. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...fellow at the Morgridge Institute for Research in Madison, Wisconsin. It is commonly cited that approximately...enzyme function, protein interaction, etc. by comparison to non-phosphorylated or ‘wild type’ control ...dephosphorylation reaction can be tracked through the comparison of phospho:non-phospho species on PhosTag gels...fellow at the Morgridge Institute for Research in Madison, Wisconsin. She is particularly interested in the...
  49. Troubleshooting and Optimizing a Western Blot

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    ...nuclear- or DNA-binding proteins, you may need to sonicate your lysates in order to release the proteins ...you’re using chemical blockers, you can run a comparison of different formulations instead of concentrations...temperature, and/or buffer to increase lysis Add sonication step to protein lysis Splice variants Check...lysing buffer/adjust lysing buffer strength Sonication based lysing protocol Subcellular fractional...  Rachel Leeson...
Showing: 201 - 250 of 718 results