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TypeBlog PostPublishedMay 20, 2021, 1:15 p.m....months. Jennifer and I spent about an hour that morning scrambling. Everything we had set up involved multiple...adapted for video making. Cycle One: The Ironing Board Control Center With the pandemic in full ...that included iPad and her laptop raised on an ironing board, giving it the perfect height (Fig. 2). We...to think of something else. Figure 2: Ironing boards. Good as a WFH desk, and as a video control...smooth filming in the future, but we were finally refining the process down to a place I could be happy with...
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10 Steps to a Perfect Science Talk
TypeBlog PostPublishedAug. 23, 2016, 2:30 p.m....iBiology which hosts an amazing amount of useful, entertaining science lab, career, and policy content. Watch...presentation appropriately. Find out who will be listening. Are they early, mid, or late career? Will many...force it. You can always relate to scientists by explaining how your work fits in the bigger science picture...that this might sound condescending, consider explaining the term on a slide but not reading the explanation...practice. I always take a poll of scientists in training to see how many in my audiences have given a 50... -
Using Phosphoserine to Study Protein Phosphorylation
TypeBlog PostPublishedJune 23, 2016, 2:30 p.m....pSer-incorporated proteins requires a molecular toolkit containing the OTS, an appropriate bacterial host strain...The pCRT7 NT TOPO vector contains a mutant pSer-containing GFP that can be used as a positive control during... researchers can run samples on SDS-PAGE gels containing Phos-Tag, a small molecule that binds specifically...that wasn’t originally a serine (e.g. mutating a threonine residue to a phosphoserine) to control for effects...large proteins by Western blotting using Phos-tag containing gel." Nat. Protoc. doi 10 (2008). Journal. Resources... -
Easi-CRISPR: Generating Knock-In and Conditional Mouse Models
TypeBlog PostPublishedApril 5, 2018, 12:42 p.m....knockout mouse in about 3 months. Image courtesy of Wenning Qin. Single-stranded DNA repair templates Short...electroporation methods using in mouse transgenics. Designing an Easi-CRISPR knock-in allele To design a knock-in...make the first two bases of the left homology arm guanine (i.e. GG); this design has been shown to increase..., you can add a silent PAM or guide mutation. Designing an Easi-CRISPR conditional allele The design for... can amplify the desired region using primers containing the homology arms on both sides and the T7 promoter... -
CRISPR 101: Off-Target Effects
TypeBlog PostPublishedFeb. 8, 2024, 2:15 p.m....Off-target effects” a vague catch-all term for unintended consequences in an experiment. For CRISPR work...In other words, they can sometimes localize to unintended sites with sequence similarity to the on-target...on-target site. When a CRISPR-Cas system localizes to unintended sites and performs its programmed function there...not Homologous Recombination or Alternative End Joining, two alternative pathways. See (Guo et al, 2023...factor, etc.) this control is usually not very meaningful. Unfortunately, there really aren’t any effective... -
Building Global Connections with the International Mentorship Program USA-EUROPE
TypeBlog PostPublishedNov. 6, 2015, 3:30 p.m.... aim of improving professional development and training in a globalized world where individuals can exchange...IMP 2014-15, University of Valencia): “At the beginning, I registered at IMP to have an opportunity to...throughout the entire year. In addition to their direct training role as mentors to Spanish mentees, mentors also...addition to a pre-program self-assessment survey defining expectations of the program and their baseline...resources, and entrepreneurial visions thereby opening new frontiers for young talent around the world... -
Tips for Working in Industry & Success in Management: Interview with Crystal Shih
TypeBlog PostPublishedOct. 5, 2017, 1:13 p.m....Crystal developed her management skills - training, learning from peers, and being receptive to those ...experiment. Chapter 6: Advice to scientists who are planning on working in industry. The importance of recognizing...our Mentoring for Scientists Series Learn about Turning Your PhD into a Career with Joanne Kamens' AMA ... -
Sequencing Options for CRISPR Genotyping
TypeBlog PostPublishedOct. 4, 2016, 2:30 p.m....thresholds for validating CRISPR experiments, many are turning to next-generation sequencing options over older...configuration, produces many false negatives Combining sequencing techniques can ensure validated experiments...Editing Specificity” by Tycko et al. 2016 and “Defining and improving the genome-wide specificities of.... Bell, Charles C., et al. "A high-throughput screening strategy for detecting CRISPR-Cas9 induced mutations...PMC3651036. 15. Tsai, Shengdar Q., and J. Keith Joung. "Defining and improving the genome-wide specificities of... -
Important Considerations When Using AAVs
TypeBlog PostPublishedJune 13, 2017, 2:30 p.m....Location, Location! Failure to Plan (for Storage) Is Planning to Fail Patience Is Bitter but Its Fruit Is Sweet...types not within the vicinity of the injection. Examining serotypes 1, 8 and 9, not only do they undergo...transport issues. 2. Failure to plan (for storage) Is planning to fail Freeze/Thaw Cycles Of course, one cannot...at the University of Manitoba. Her project is examining the use of DREADDs (Designer Receptors Exclusively...Activated By Designer Drugs) to determine the role of serotonin in movement. Follow her on twitter @katrinaboards... -
Make a Splash: Notions of Scientific Impact Are Evolving
TypeBlog PostPublishedDec. 5, 2014, 2:54 p.m....articles in less-prestigious journals have been gaining ground. A study issued by Google in October entitled...report also noted, papers detailing Nobel Prize-winning research typically weren’t among the most cited...impact.” As we look ahead to the end of a year and beginning of another, perhaps it’s worth reflecting on a...Other Novel Measures for Scientific Impact" from Opening Science - The Book Bollen J, et al. "A Principal... -
2022 and 2023: Reflections and Looking Forward
TypeBlog PostPublishedFeb. 23, 2023, 2:15 p.m....countries. We, like everyone around the world, are learning to navigate the changes brought about by the pandemic.... As always, we seek to understand our role in opening up science to help create new knowledge and research...opportunities. I have spent the last 9 months quietly learning about Addgene’s past and present, but the real...place in it. This period of careful reflection and planning will allow us to continue to serve the scientific... -
Addgene's Top Blog Posts from 2020
TypeBlog PostPublishedDec. 29, 2020, 2:15 p.m....providing cloning grade DNA to you, we'd save you days of time so that you can begin your cloning experiment...this year were influenced by COVID-19 and our new cloning grade DNA service. We've had 11 blog posts on COVID... and industry. This year, we also launched the cloning grade DNA service. This service features 210 plasmids... -
Science Communication Snapshot: DayCon 2015
TypeBlog PostPublishedJune 17, 2015, 9:04 p.m....was fortunate to see the event planning unfold from the very beginning. The board picked a theme (Bio-...DayCon DayCon and other similar events show how meaningful science outreach can be, both for scientists ...communication and organizational skills provide great training for a number of alternative careers in science... -
Antibodies 101: Introduction to Antibodies
TypeBlog PostPublishedJan. 19, 2021, 2:15 p.m....the hybridoma can occur, known as genetic drift, meaning that over time, the antibody you expect it to produce... the advantage of being significantly smaller, meaning they can recognize epitopes that full antibodies... detect the presence of proteins from samples containing a mixture of proteins. The proteins are separated...antibodies to detect proteins is referred to as staining. This panel of antibodies is used to both identify...enzymatic reaction. Figure 5: Immunofluorescence staining for CD31 (green), CARD14 (red), and pNFkB (blue... -
SciComm with the Experts at Science in the News Part 2
TypeBlog PostPublishedApril 27, 2017, 2:30 p.m....Chapter 10: Maintaining Institutional Knolwedge and Continuity - SITN's key technique for retaining institutional... topics discussed during the interview. Happy listening! Listen to the interview with Science in the News...communicating science to the general public and training the next generation of science communicators. ... -
Celebrating Accomplishments in the Lab
TypeBlog PostPublishedMay 8, 2014, 3:07 p.m.... She suggests to pick up hobbies (for her its gardening) that help you relax and enjoy life. “I love academics... the afternoon during the child-rearing years, learning “lots of interesting things from children, their... cooks and cleans with his family, “sometimes listening to audiobooks.” Church and his family spend family...Advice for Choosing a Research Project Finding and Joining Your Dream Lab Advice for Choosing a Research Project... -
"What Makes a Good Mentor?" and 6 More FAQs About Science Mentoring
TypeBlog PostPublishedDec. 17, 2013, 2:47 p.m....scientists. It takes us a decade or more for just the “training” portion of our careers. In that time we may have...organizing mentoring programs for scientists and doing training to help mentors and mentees have fulfilling, productive...these things and more. It is instructive (and entertaining) to review this list of bad mentor behaviors...openly Gives honest feedback without ever being demeaning Does not micromanage (teaches you to fish, doesn... -
Tag Your Favorite Yeast Genes with Ease
TypeBlog PostPublishedNov. 19, 2013, 2:37 p.m....surprisingly simple, requiring only a PCR product containing the modifying sequence flanked by approximately...distributes several ready-to-use, modular plasmids, combining fluorescent tags, epitope tags, protease sites...Formosa has deposited six plasmids with a multiple cloning site in place of the epitope tag for creation of...additional benefit of multiple selection markers for combining modifications within a single strain. In addition... -
Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies
TypeBlog PostPublishedNov. 27, 2019, 2:03 p.m....Understanding the biology behind brain development, learning, and behavior poses many challenges in neuroscience...are the three main takeaways from the paper for designing experiments with AAV: 1. Getting your viral vector...defined neuronal populations using viral strategies Combining different approaches can increase the specificity...information. To complement these efforts, we’re beginning to share data through our recently launched AAV... -
IBEX Knowledge-Base: A Data Resource for Multiplex Tissue Imaging
TypeBlog PostPublishedJune 8, 2023, 1:15 p.m....conventional microscopes, amplified staining methods, and oligonucleotide-based staining. Of course, if you’re interested...multiplexed tissue imaging method that uses iterative staining and chemical bleaching to allow for high-resolution... inspired by the open-software community when designing the IBEX Knowledge-Base, seeking to create an ... -
Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation
TypeBlog PostPublishedMay 21, 2020, 1:15 p.m....in Drosophila. SPARC was developed by Thomas Clandinin’s lab, and incorporates several common genetic ...both for visualization and for optogenetics. The Clandinin lab used SPARC to image the calcium response in... YE, Wang IE, Ishida IG, Maimon G, Wilson RI, Clandinin TR (2019) SPARC: a method to genetically manipulate...resources on the Addgene blog Learn about Gateway cloning Read about genome engineering in flies Learn more... -
Oh, The Places You Can Go: Careers in Science Communication - Science Writing
TypeBlog PostPublishedOct. 3, 2017, 12:11 p.m....away-from-the-bench career option. So if you’re interested in learning more about science communication careers, you’...s paid off. Work is never boring. “I just keep learning on the job. It keeps it interesting.” And while...accomplishment.” He started working on them shortly after joining IDT when his boss was looking for someone to take...never get off of the ground. So limiting that, and defining a couple of key terms is a better approach. Also... -
Bringing Sustainable Practices to the Lab: Recycling
TypeBlog PostPublishedFeb. 14, 2019, 2:35 p.m....keep reading). Corning takes back and recycles a lot of their plastic packaging (from Corning®, Falcon®, ...plastics are ever recycled, so adopting a few meaningful changes in the lab could make a big difference...person! If you use serological pipettes from Corning®, Falcon®, or Axygen®, set up a bin or box in your... -
Addgene's a Nonprofit? Nonprofit Awareness Day 2017
TypeBlog PostPublishedJune 5, 2017, 3:19 p.m....make us self-sustaining. With this unique model, we are able to spend resources on maintaining quality and...investors or stock. All of our proceeds go into running the repository and serving scientists better. That... this trust by keeping excellent records and maintaining confidentiality where it is appropriate. This... -
Sensing Neuronal Dopamine
TypeBlog PostPublishedJuly 30, 2019, 12:59 p.m.... hormone,” it’s also involved in reinforcement learning, decision making, and motor control. What do... mice Report modulation of DA signaling during learning and reward consumption in live mice Find dLight...Drosophila Detect DA dynamics in response to a threatening looming stimulus in vivo in transgenic zebrafish...release in live mice over the course of Pavlovian conditioning Find the GRABDA sensor here! Overall these ... -
An Addgene Co-op: The Intersection of Fun, Friendship, and Work
TypeBlog PostPublishedJune 29, 2023, 7 p.m....connections with coworkers and doing something meaningful with my career. I can’t pinpoint exactly when... mine is mainly within content and business. Joining the Addgene crew! This pattern was consistent:...bigger projects I was tasked with was creating and sustaining a biweekly report of our social media analytics...free time, she enjoys reading, petting dogs, and winning the office Just Dance competition. Special ... -
Celebrate Open Data Day with Addgene’s AAV Data Hub!
TypeBlog PostPublishedMarch 5, 2020, 2:15 p.m....when you’re running your next AAV experiment, use these questions as a guide for designing your future...Data Hub to quickly find details they need for planning their next experiments using a specific AAV tool...activate receptors? If you’re in the process of designing your experiment, read our blog post on important... -
Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life
TypeBlog PostPublishedApril 12, 2018, 1:02 p.m....instruction. Six of the kits, known collectively as Greening the Classroom, were developed and tested by ABRC...such as instructional videos and datasheets. The remaining kits were donated to ABRC by the Arabidopsis research...has many benefits. Engaging students in active learning through hands-on experiments allows them to develop... out opportunities to engage people in science learning. Whether visiting classrooms, hosting field trips... -
Identifying Sequence Elements with SnapGene's Feature Database
TypeBlog PostPublishedDec. 21, 2017, 2:06 p.m....daily basis, many found these tools inadequate for planning, visualizing, and documenting their procedures...procedures. Preventable errors in the design of cloning strategies set experiments back days or even weeks. Primer... even if one or two codons are missing at the beginning or end of the feature. With the increasing popularity...people love to use, and likes to start at the beginning, shepherding a new product from the idea stage... -
The Strength of Story Telling
TypeBlog PostPublishedDec. 19, 2017, 2 p.m....to have a story structure: beginning, end; cause, effect; lesson, learning. Storytelling is a crucial ...part of interpersonal communication, which makes honing your ability to tell stories a similarly crucial... with me. First I calmed the situation down by opening a dialogue specifically about the project and contributions... -
What Do I Do Now? Academic v. Non-Academic Career Decisions
TypeBlog PostPublishedMarch 9, 2017, 3:40 p.m....acknowledged perks of scientific and technical training is that these educational paths prepare you for...perspective on other types of science careers as their training progresses. In From Academic Solos to Industrial...explicit and implicit aspects of today’s postdoctoral training can directly interfere with a seamless jump into...quickly Jack/Jane of all trades More sources of training and resources within the company Must find own... -
HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides
TypeBlog PostPublishedOct. 10, 2019, 12:41 p.m....constructed by grafting different parts together. By combining two optimized elements of an antibody probe, scientists...fused them to sequences called complementarity determining regions (CDRs, or loops) of an antibody that ...bound to HA epitopes for minutes in living cells, meaning that scientists can track the movement and localization... & Givol, D. (1972). Localization of antibody-combining sites within the variable portions of heavy and... -
5 Uncommon Ways to Gain Career Skills During an Uncommon Time
TypeBlog PostPublishedOct. 20, 2020, 1:15 p.m....out-of-the-box ideas that might jump start your learning. 1. Create a peer mentoring group We don’t all...mentors to listen, guide and support us during training and throughout our careers. An easy way to access...academia and not in academia is run by grant funding. Learning to write grants quickly, efficiently and effectively...Funding For Your Research University of Florida: Training Grants and Other Funding Opportunities L’oreal... -
Summer SciComm Series: Master of SciComm
TypeBlog PostPublishedJuly 21, 2022, 1:15 p.m....that my piecemeal degree might one day foster a meaningful day job only came into focus when an English ...podcasting, philosophy of science, creative writing, designing and curating exhibitions, animation, graphic design...interested in connecting with the public in a meaningful way, and not solely to ensure a profit. Often...Cognitive LoadSummer SciComm Series: Modes of CommunicationIntro to Our Summer SciComm Series!... -
Three Tips for Preventing Viral Plasmid Recombination in Your Samples
TypeBlog PostPublishedSept. 14, 2021, 1:15 p.m....the transgene will be lost. Over time, bacteria containing the recombined vector backbone will also outgrow...samples. For a more detailed comparison of these cloning strains, browse the table of common gene mutations...there are both large and small colonies. Bacteria containing the recombined product tend to grow faster, so...N, Huo Y-X (2020) Reversed paired-gRNA plasmid cloning strategy for efficient genome editing in Escherichia... -
Even more elegant: Single injection CRISPR/Cas9 in C. elegans
TypeBlog PostPublishedJuly 7, 2015, 3:36 p.m....have used PCR-based screening to detect mutants, but this method limits screening to a few hundred worms...In the summer of 2013, a remarkable nine papers describing CRISPR/Cas9 genome engineering methods for...background. Their new method uses a selection cassette containing genes for hygromycin resistance and the Roller... -
Extracurricular Activities for a Strong Science Career Path
TypeBlog PostPublishedJan. 21, 2014, 3:08 p.m....Science Careers has developed a web-based, career-planning platform called the Individual Development Plan...have to be ‘extra’ Today’s graduate and postdoc training focuses on using multidisciplinary, collaborative...incorporating EAs into your routine. Getting started: Planning your career path and identifying needed skills...suit these skills and interests: Use career planning tools such as the Individual Development Plan ... -
Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs
TypeBlog PostPublishedDec. 20, 2017, 2:33 p.m.... If you are in this position, you should start planning for your visa and or green card process well before...about the process early, keeping good records, and planning for the route to a rewarding career. It is more...this post, we discuss the following paths to maintaining a status in the United States including green... for self-sponsorship of the I-140 application meaning that the category does not require a foreign national... your current PI and Ph.D. or Masters PI. The remaining letters should be from independent people – i.... -
Plasmids 101: NGS Quality Control for Pooled Libraries
TypeBlog PostPublishedOct. 26, 2017, 1:59 p.m....Addgene also distributes pooled plasmid libraries containing hundreds, thousands, or even a million plasmids...plasmid. However, lots of library DNA is needed for screening experiments, so the scale of the DNA prep is larger...where each transformant forms its own colony, uninfluenced by the growth of other cells. For libraries ...wishes you luck in your future library use and screening, and if you have any tips, input, or stories of... -
Visualizing Genomic Loci with CRISPR-Sirius
TypeBlog PostPublishedApril 18, 2019, 12:53 p.m....result in a blue, green, or red readout. But gRNAs containing two different hairpins will produce a secondary... depending on the hairpin combination. A gRNA containing all three hairpins produces a white color. While...simultaneously, Ma and Pederson created another sgRNA containing the PP7 hairpin sequence (sgRNA-Sirius-8XPP7)...CRISPR-Sirius. Triple color detection With a third sgRNA containing alternating MS2 and PP7 sequences (sgRNA-Sirius... -
How Dry Ice Affects Viral Vector Sample pH & How to Avoid It
TypeBlog PostPublishedJune 20, 2017, 2:30 p.m....prevented. Keep this information in mind if you’re planning on shipping something on dry ice or if you’re ...they would be exposed to a closed environment containing CO2 (from the dry ice). After 48 hours of CO2...it back in the freezer? We then tested whether returning the tubes to the -80°C freezer after storage on...Murphy BM, Swarts S, Mueller BM, van der Geer P, Manning MC, Fitchmun MI. Protein instability following ... -
The MTA at Addgene
TypeBlog PostPublishedJuly 23, 2024, 2:17 p.m.... in our repository. Instead, we facilitate the signing of MTAs by the Provider and Recipient (in Addgene...under which your material will be distributed. In signing this agreement, your organization agrees to let... a form needs to be signed. But the truth is, meaningful progress depends on things being organized, tracked... be used. Setting clear expectations from the beginning makes the sharing process smoother, more consistent... -
Predicting Adverse Reactions to Monoclonal Antibody Drugs
TypeBlog PostPublishedFeb. 13, 2024, 2:15 p.m....Antibodies administered as drugs are still immunogenic, meaning that they elicit an immune response from the body...the safety and the efficacy. Better together Combining in silico and in vitro approaches enables researchers...method of immunogenicity assessment can help in determining the T-cell epitopes that will likely bring about...silico Immunogenicity Assessment for Sequences Containing Unnatural Amino Acids: A Method Using Existing... -
K. phaffii: Rising to the Occasion
TypeBlog PostPublishedApril 4, 2023, 1 p.m....obligate aerobe. It’s also a methylotropic yeast, meaning it can utilize methanol as its sole source of carbon... that it predominantly uses non-homologous end joining (NHEJ) to perform DNA double strand break repair...Organisms Plasmids 101: Yeast Vectors Tips for Screening with the Yeast Two Hybrid System Additional...Additional Resources on Addgene.org Yeast-mediated cloning CRISPR plasmids: Yeast Yeast Synthetic Biology... -
Multiplexed Overexpression of Regulatory Factors (MORF) collection
TypeBlog PostPublishedMarch 30, 2023, 1:15 p.m....) act as the gate keepers for gene expression, turning transcription on and off by binding proximal to... library can be used with other selection and screening mechanisms, although the unique barcodes for each...mutation and/or TF dysregulation. A targeted TF screening approach can not only lead to a better understanding...Cas-9 knockout and transcriptional activation screening. Nat Protoc. 2017 (4):828-863. doi: 10.1038/nprot... -
Components of CRISPR/Cas9
TypeBlog PostPublishedFeb. 2, 2016, 5 p.m....Campylobacter jejuni (CJ) 3' NNNNRYAC Neisseria meningitidis (NM) 3' NNNNGATT Streptococcus thermophilus...their target sequence of choice (see tips for designing a gRNA) and their prefered Cas9 variant (with ...bacterial genomes are transcribed into pre-crRNAs containing both the spacer region and the direct repeat ...thinking about the types of experiments you're planning on doing and read the eBook or browse our CRISPR... -
Addgene Welcomes John Doench to the Advisory Board
TypeBlog PostPublishedAug. 13, 2015, 2:30 p.m....everything that could be done with RNAi in terms of determining what genes are involved in biological process...the real hits is so much shorter…. it’s a great screening technology.” Doench thinks that the future of...become more efficient in terms of editing and screening) such that thousands of individual edits can be...says he’s “known of Addgene basically since the beginning and [has] been using Addgene ever since” and that... -
Resume Writing for Non-academic Science Careers
TypeBlog PostPublishedDec. 18, 2014, 4:06 p.m....of the University of British Columbia. When transitioning from an academic science career path to a non-academic... lectures? These transferable skills could be meaningful for your potential employer. For example, my ... Developing Transferable Skills During Science Training and So You Think You have Skills. Get Science...graduate students in job searching. Start your career planning early, start now Don't wait until the day you ... -
Fluorescent Proteins 101: Luciferase
TypeBlog PostPublishedJune 24, 2014, 3:59 p.m....Shortly after the initial article describing the cloning of firefly luciferase was published in 1985, several...Luciferase reporter assay The typical luciferase-containing plasmid is most often used to investigate the...cloned into a plasmid downstream of luciferase containing a stop codon, which is then transfected into ...into the desired cells. A hybrid mRNA transcript containing luciferase and the gene of interest permits translation... -
New and Upcoming Viral Vectors - May 2020
TypeBlog PostPublishedMay 28, 2020, 1:15 p.m....Since the beginning of our viral service in 2016, we’ve added many new tools to our inventory of ready-to-use...plasmids expressing SARS-CoV-2 proteins at the beginning of April. Now, we've turned some of the most requested... genes and is used for genome-wide inhibition screening. Find the Dolcetto lentiviral prep here Control...Fishell Chemogenetics Chemogenetics offers noninvasive methods for manipulating cell activity, particularly...