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  1. Editor's Choice, September 2016

    Type
    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...

  2. 28 Hot Plasmid Technologies from 2015

    Type
    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...

  3. Bricking Science: Portraying Scientific Reality Through LEGO

    Type
    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...

  4. Management for Scientists: Giving Feedback

    Type
    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...

  5. Choosing a Good Mentor for Scientists

    Type
    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...

  6. The Strength of Story Telling

    Type
    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...

  7. What Do I Do Now? Academic v. Non-Academic Career Decisions

    Type
    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...

  8. Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring

    Type
    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...

  9. Another Pathway into Cells: iTOP

    Type
    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...

  10. Antibody Validation for Flow Cytometry

    Type
    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...

  11. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    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 ...

  12. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    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 ...

  13. Addgene Depositors Get More Citations

    Type
    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...

  14. K. phaffii: Rising to the Occasion

    Type
    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 (...

  15. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    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...

  16. Four Factors that Differentiate the Stem Cell Field

    Type
    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...

  17. Synthetic Photobiology: Optogenetics for E. coli

    Type
    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...

  18. Management for Scientists: Delegating is Key

    Type
    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...

  19. Sweating the Small Stuff: Details in the Lab

    Type
    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...

  20. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    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...

  21. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...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...

  22. Fluorescent Proteins 101: Introduction to FRET

    Type
    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...

  23. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    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...

  24. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    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...

  25. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...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...

  26. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...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...

  27. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...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...

  29. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ... 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)...

  30. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  32. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...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...

  33. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...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...

  34. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...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...

  35. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...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 ...

  36. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ...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 ...

  37. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...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...

  38. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...progeny are produced (Figure 1).  Figure 1: Comparison of the life cycle of a lentiviral vector with...integrase-deficient lentiviral vectors? There are several reasons why you might choose an IDLV over a lentiviral ...This blog post was written by Addgenie Rebecca Hutcheson.   References and resources References Banasik...PMC2835240 Wu Y. (2004). HIV-1 gene expression: lessons from provirus and non-integrated DNA. Retrovirology...

  39. March for Science

    Type
    Blog Post
    ...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...

  40. Tips for Getting a Faculty Position

    Type
    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...

  41. Using Phosphoserine to Study Protein Phosphorylation

    Type
    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...

  42. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...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...

  43. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...Addgene. Klann TS, Black JB, Chellappan M, Safi A, Song L, Hilton IB, Crawford GE, Reddy TE, Gersbach CA...publication at Addgene. Maeder ML, Angstman JF, Richardson ME, Linder SJ, Cascio VM, Tsai SQ, Ho QH, Sander...Sander JD, Reyon D, Bernstein BE, Costello JF, Wilkinson MF, Joung JK (2013) Targeted DNA demethylation... this publication at Addgene. Mendenhall EM, Williamson KE, Reyon D, Zou JY, Ram O, Joung JK, Bernstein...

  44. CRISPR Guide

    Type
    Guide
    ... A., Chan, M. M., Bauer, D. E., Marson, A., Parsons, L. R., & Adamson, B. (2024). Improving prime editing...including transposons, integrases, and recombinases, with Cas enzymes. CRISPR Transposases Transposon systems...I., Yoon, Y., Song, C., Cao, Y., Gallant, J., Xue, W., Rivera-Pérez, J. A., & Sontheimer, E. J. (2019)...S., Cofsky, J. C., Kranzusch, P. J., Sontheimer, E. J., Davidson, A. R., Maxwell, K. L., & Doudna, J. ..., J., Edraki, A., Shah, M., Sontheimer, E. J., Maxwell, K. L., & Davidson, A. R. (2016). Naturally occurring... Chen, C., Nelson, J. W., Newby, G. A., Sahin, M., Osborn, M. J., Weissman, J. S., Adamson, B., & Liu,...Ramadoss, G. N., Shi, Q., Hung, K. L., Samelson, A. J., Pogson, A. N., Kim, J. Y., Chung, A., Leonetti...

  45. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ... appear within a range of wavelengths. For that reason, we speak of excitation and emission spectra. Both... happens due to a process called Fluorescence Resonance Energy Transfer (FRET).   Figure 1: Emission...fused to CD45, but is separately expressed. (The reason for having a separately expressed transduction ...difficult. I chose BV421 for the following two reasons: 1) The emission spectrum of BV421 is far enough...

  46. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ... work, funding for science, and numerous other reasons. It is important to remember that we, scientists...Albany, NY, USA 42.6526°N 73.7562°W – Nicholas Thompson Over the last few years, there's been a disturbing...positive event that I hope happens again. Nicholas Thompson is a PhD Candidate studying Nuclear Science and...enforcement, and, although there was at least one person vocally protesting us, he was not threatening in...

  47. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...Single-chain variable fragment (ScFv) Figure 2: Comparison between the IgG antibody and scFv. Because...Variables (scFvs). Nanobodies Figure 3: Comparison of the Hcab and nanobody. Nanobodies are the...10.1016/j.ymeth.2017.01.009 Crivianu-Gaita V, Thompson M (2016) Aptamers, antibody scFv, and antibody...antibody Fab’ fragments: An overview and comparison of three of the most versatile biosensor biorecognition ...

  48. Which Fluorescence Microscopy Technique is Best for Me?

    Type
    Blog Post
    ...Many thanks to our guest blogger, Doug Richardson! Doug Richardson is the Director of the Harvard Center...This post was contributed by Doug Richardson, Director of the Harvard Center for Biological Imaging and... 2, 932-940 (2005). PubMed PMID: 16299478. 9. Richardson DS, Lichtman JW. Clarifying Tissue Clearing. ...

  49. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ... something is working for you, then there is no reason to change it. But what if a kit isn’t working for...variety of manufacturers. It also includes a rough comparison of list price between these manufacturers, as...kits before deciding which one to buy. Table 3: Comparison of popular plasmid purification kit options ...blog post was written by Ashley Waldron, Hannah Dotson, and Nyla Naim. Resources Additional resources...

  50. #DeckTheLab with Tinsel and Test Tubes, Fa La La La La La La La La

    Type
    Blog Post
    ...for near-freezing temperatures, our first of the season snow flurries, dusting off the winter coats and... to #DeckTheLab with virus snowflakes from @CVRHutchinson #SciArt #STEAM #WinterWonderlab @Addgene Make...WFH space feel a little more festive this holiday season! It is that time of the year again: miniature ...
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