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...reproducible assay. That information can make you a much stronger candidate!  Additional uses The cover letter is...job with a lot of file management that asks for strong organizational skills. You might think, “I’d be...letter The opening paragraph is the place to introduce yourself and your background. It’s a great place...and is important for the job]. Notice that the intros above allow you to highlight what specific skills...

... Martin et al., 2018). The ACE reporter is a bicistronic construct that consists of a mutated mCherry ...eGFP. To create the inactive mCherry, the lab introduced a 43 base-pair insertion that disrupts fluorescence...use across different cell types and systems. To introduce flexibility, the Dow lab created a base editing... optimized cytidine base editor. The lab then introduced an sgRNA targeting the GFPACG to the cells. As...

...founder of Research Space. There are many types of electronic lab notebooks (ELNs), each with its own pros ... virtue of liberating data from paper into an electronic environment and hence making it searchable and... of Research Space, which provides the RSpace electronic lab notebook. Rory is based in Cambridge, Massachusetts...on the Addgene blog Find tips on choosing an electronic lab notebook Watch a livestream of the Minisymposium...

...protocols therefore differ from library to library. We strongly recommend following the protocols provided by ...a low multiplicity of infection (MOI). MOI is controlled by viral titer, so you’ll need to measure your...Addgene uses a physical titer (via ddPCR) to quality control our viral preps, we recommend determining the functional...transduction protocol you can follow and a troubleshooting blog should you run into any issues.  Keep ...

...and selection for targets can be done largely in vitro; Easy and reproducible commercial synthesis (those..., Akhtar and Marty, Jean L. “Aptamer based electrochemical sensors for emerging environmental pollutants...systems using immobilized aptamers.” Biosens Bioelectron. 2011 May 15;26(9):3725-36. PubMed PMID: 21419619...FRET in Your Experiments Learn more about GFP Controlling Protein Activity with GFP Resources at Addgene.org...

...social sciences go uncited. Creative titles and controversial topics can garner some attention, though in ...Journal of Number Theory (1,519 views), and Gastroenterology (1,046 views) rounded out the top four average...infections. The video abstracts provide a brief introduction to the research and, after watching the video...links to the videos, keeping the copyright and control over the videos with the researcher and the host...

...overexpression vector rather than going through all the troubleshooting required for Cas9-based activation. CRISPRa...Dow D, Buser-Doepner C, Bertotti A, Trusolino L, Stronach EA, Saez-Rodriguez J, Yusa K, Garnett MJ (2019...Weissman JS (2014) Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Cell 159:647...2015) A light-inducible CRISPR-Cas9 system for control of endogenous gene activation. Nat Chem Biol 11...

...reviewer’s concerns, you are making your hypothesis stronger. When you share data at a conference poster session... can frequently be traced back to one of a few trouble spots:  Communication (or lack thereof): Information...Empathizing with people, especially people with whom you strongly disagree, is a critical component of conflict ...tools I’m mentioning here, this may be the most controversial, so I would like to qualify it. If you are attempting...

...Level 1 microbe makes it a perfect candidate for introductory teaching and research environments (Idalia &...study human cardiovascular disease, specifically stroke and hypertension (Iannaccone et al., 2009). Rodents...Resource, a breeding facility for X. laevis and X. tropicalis, the two most commonly used species of frog. ... study human disease-relevant genes in such a controlled environment displays how essential the frog is...

...while shRNA and siRNA are synthetic constructs introduced through external methods. shRNA processing Following...expressed constitutively, either in lentiviral or retroviral backbones. On some occasions, shRNAs are transiently...transient transfections and are handy for quick in vitro experiments. shRNA plasmids typically use RNA polymerase...common promoters are U6, H1, and 7SK. These are strong promoters with no tissue specificity, so in some...

...neither the aptamer nor the small molecule are strongly fluorescent but the binding of an aptamer to its...plasmids encoding aptamer-tagged RNA and later introducing the small molecule into the cell. The fluorescence...metabolites, leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches...then identified through microfluidic-assisted in vitro compartmentalisation for their high binding affinity...

...most important ways to spatially and temporally control gene expression, especially in in vivo models, ...Golgi staining, injection of diffusible labels, or electron microscopy of stained sections, were limited due...functionally similar to Cre-based Brainbow, but is controlled by FLP/FRT recombinase. When placed under different.... The use of AAV provides spatial and temporal control without the need for germline modification, and...

...priorities and needs, but sometimes it just takes introspective thought on what’s worked for you and not worked...reputation? If you are choosing a training lab, I strongly suggest you complete a work rotation in the lab... it could also be a sign that the lab head has trouble attracting scientists. To get a better idea of ...and non-academic career paths. I can’t say this strongly enough - don’t choose a mentor that sees you only...

...formation in the genome and are inefficient at introducing point mutations. In 2016, the researchers at ... Broad Institute envisioned a new strategy to introduce point mutations in the genome. They fused a catalytically...base editors (CBE) use a cytidine deaminase to introduce a C→T change at precise locations in the genome...his super giant albino gorilla fighting off a monstrous wolf and crocodile, that were “affected by CRISPR...

...as the insertion of a fluorescent tag or the introduction of a specific mutation, one generally relies...one but two rounds of single cell cloning – as a control, one should revert the edit back to the original...technologies offer an alternative to HDR for introducing edits. The same locational constraints are even...the method of producing the guide (synthetic, in vitro transcription, or lentiviral delivery) can affect...

...researcher in the University of Michigan Department of Astronomy. She studies how planets form around young stars...community of science supporters is much larger and stronger that I expected. Jennifer Stancill, Ph.D., is ... an agenda that halts current data collection, destroys existing data, and guts future funding.I marched...march read "You know things are serious when the introverts arrive." I couldn’t watch the denial of facts...

...Correlative light and electron microscopy (CLEM) is an imaging technique that combines electron microscopy (EM...cellular structures (0.5-1% osmium tetroxide). These conditions destroy most PA-FPs, but, in 2015, the Looger...

...initiate our MTA process and perform standard quality control but will not make the plasmids available until:...Continue to Next Step". As part of our quality control process, we will sequence the full plasmid and ...recombination in bacteria) such as lentiviral, retroviral, and AAV plasmids, we recommend the NEB Stable...received them and they will undergo our quality control process to verify your plasmids. Our friendly Addgene...

... Using the FITS Approach Caitlin Faulds, who introduced the very first antibody deposit (the Integrin...flow cytometry expertise in three posts. An Introduction to Gating in Flow Cytometry, Designing Your ...two posts. Flow Compensation and Flow Cytometry Controls Priyamvada Prathima, who wrote a helpful guide...

...Podcast breakdown (Time : Topic) 0:00 - 2:00 : Introduction to the podcast from producers Tyler Ford and...and Eddy Page. 2:00 - 3:55 : Alex introduces himself and explains why he decided to do an MD PhD - it's...in generating Cas9 based tools to enable facile control of DNA and RNA.     Special thanks to our sound...

...ultimate prize – a glorious trophy. Sure it’s plastic and you have to share 1 trophy among 6 people, but you...occurring earlier but that the photo evidence was destroyed to protect the innocent revelers. Our Halloween...

...interview with Addgene's founders Podcast breakdown Intro Chapter 1 - Who are Addgene's Founders? Chapter ...the importance of sharing information/knowledge Outro Many thanks to our guests, Melina Fan, Kenneth ...nonprofit? Get an Inside Look at Addgene's NGS Quality Control Addgene's Viral Service - Why Virus? Why Now? ...

...AAV9 tdTomato virus in the mPFC.  The AAV9 is so strong that you can even see dendritic spines!  Curious...Images that clearly show viral spread and cellular tropism* (min. 300 dpi, PNG format) At least two replicates... the target region, if the report describes a retrograde serotype An abstract that summarizes your experiment...

...grant to order plasmids she can use as positive controls in PCR-based cervical cancer screens. This third...predominantly HPV 16 and 18.” HPV plasmids as positive controls To start her work, Gladys will improve cervical... containing a variety of HPV genes as positive controls in her PCR screens. She hopes that these screens...

...projects 0:28 - 1:12 : Introduction from Tyler 1:13 - 2:02 : Niroshi and Ben introduce themselves 2:03 - 3...your biggest experimental success? 31:32 - end : Outro In our next episode, Niroshi and Ben discuss how...

...parental cell line, but not in the iso-genic knockout control. This can involve cell surface labeling and/or ... Additionally, it can be a real challenge to troubleshoot RNAi failures. Potential explanations for RNAi... target of interest, you can add an additional control containing treated cells. The expected change in...55133-7 More resources from the Addgene blog Introduction to Gating in Flow Cytometry Antibodies 101: ...

...the performance of different nucleases. Nicola Patron’s lab compared the efficiency and specificity of...plasmids Anti-CRISPR proteins for optogenetic control of CRISPR-Cas9 Anti-CRISPR proteins that inhibit...has adapted anti-CRISPR proteins for optogenetic control of CRISPR-Cas9. By combining AcrIIA4, a Streptococcus...

...They kicked off their day with breakfast and an introduction to Addgene with our Executive Director, Joanne...in-depth look at some of the teams at Addgene. An introduction to plasmids with Director of Biology Lianna ...our UK team member Benoit Giquel. Bluegene’s globetrotting impressed our guests, as did how much work is...

...for use as a negative control, and empty backbones such as pBABE-puro for retroviral expression and pLVTHM...psPAX2. Actually, these two plasmids from Didier Trono's lab are our two most popular plasmids of the past...

... they do so by activating signaling pathways controlling properties like growth, survival, and differentiation...inactivated the pathway. “The idea is that if you introduce them into cells one at a time, you could identify... for pathways that drive resistance to drugs in vitro, and they’ve shown that it works. But, he says that...

...growth based on data voluntarily made available by Medtronic via The Yale University Open Data Access (YODA...got a nine percent bump in citations for their trouble, even when all kinds of other factors, including...universities. We particularly recommend Eric Lander's intro to biology course "The Secret of Life." 10)     ...

...for packaging: AAV1, AAV2, AAV5, AAV8, or AAV Retrograde (AAVrg) capsids.  Please be aware that this ordering...Packaged on Request prep and run the same quality control assays we run with all of our in-stock viral preps...plasmids packaged in AAV1, AAV5, AAV8, and AAV Retrograde to be 1x1013 and titers for AAV2 to be 7x1012...

...McClean’s lab generated an optogenetic toolkit that controls repression of gene expression in Saccharomyces...transcriptional repressor Mxi1, and an optogenetic tool that controls nuclear localization (LINuS). Blue light renders...mRNA decreased after 20 minutes in xylose (which controls expression of dCas9). The lab demonstrated the...

... system is toxic to proliferating cells. A Cre-estrogen receptor ligand binding domain-fusion can prevent... toxicity in proliferating Drosophila cells by estrogen-dependent activity regulation. Heidmann D, Lehner... BAC/PAC recombineering techniques. Semprini S, Troup TJ, Kotelevtseva N, King K, Davis JR, Mullins LJ...

... me like the postdocs are taking some steps to control their own future in science (see my previous blog...vehemently about the need for change.  Dr. Bourne strongly believes this is an amazing time of promise for... Novartis industry postdoctoral program.  His strongest comments were about the astounding lack of reproducibility...

...Getting into the office for the first time, being introduced to all the members around the office one by one...and undigested samples of the clones on an electrophoresis gel in order to differentiate which clone had...Once we had gotten our results back from the electrophoresis, my manager and I were unsure if there were...

...and/or the use of chelating agents that can be introduced in the purification protocol to remove endonucleases... your preparation for a long time. Tris buffer controls the pH, while the EDTA chelates any divalent cations...of using a Tris buffer is that it allows better control of the pH preparation stabilizing the plasmid, ...

...the workforce, but my main concerns were building strong connections with coworkers and doing something ...interested in knowing you as a person, builds a strong sense of trust between coworkers. Since I trusted...about making connections with coworkers seem, in retrospect, nonsensical. I have found the people at Addgene...

... sections of a paper, the abstract, figures, introduction, etc. so as to thoroughly explain the do’s and...reader is not in their head. The scientist has to introduce the reader to the manuscript and engage them in...to make the final product with an interesting introduction and concise discussion. Many thanks to our ...

...sequence the plasmid to confirm no mutations were introduced during PCR or any subsequent steps. Using high...DNA polymerases, I have never had any mutations introduced into my inserts. However, with any PCR-based ...PCR-based cloning technique there is a chance of introducing an unwanted mutation. Tips for high efficiency cloning...

...recognition come from the successes of the team. A strong sense of engagement in the work Behind-the-scenes...    This is the first in a 5 part series on Introduction to Managing People for Scientists. Subscribe...follow the entire series as it is published. Good introductory reference books on managing people: Skills ...

...function of Your Gene Of Interest (YGOI) in a controlled environment under various conditions. The first...expression system and go for a lentiviral or a retroviral backbone. This is considered to be a more robust...great for overexpressing YGOI in an isolated, controlled environment. In most cases bacterial expression...

... store a record of invading phages but to also destroy the phage upon re-exposure (2, 3). More specifically...complex binds to the foreign DNA, a cut is made to destroy the invader. Wikipedia, accessed 25 November...ultimate victor will be the species who has the strongest will to survive. Genome engineering and future...

...cases, pay special attention to designing your controls in validation, as you may need to test different...great deal of frustration when designing and troubleshooting immunoimaging assays. It can also elucidate...Antibodies 101: Polyclonal Antibodies Antibodies 101: Introduction to Antibodies Resources on Addgene.org ...

...One of the main tenets of our quality control (QC) mantra is to do our utmost to ensure that scientists...Gene Therapy. Existing methods for AAV quality control Historically, AAV vector QC has been sparse including...number of AAV preparations this level of quality control is sufficient but for viral vector core facilities...

...the origin of replication (ori) and all of its control elements. The ori is the place where DNA replication...depending on how they are regulated. Generally, control of replication is referred to as "relaxed" or "...Vectors Copy Number+ ori Incompatibility Group Control pUC ~500-700 pMB1 (derivative) A Relaxed pBR322...

...researchers and educators in 60 countries each year. Introducing Arabidopsis Arabidopsis is an important model...develop a new teaching resource or you want to introduce Arabidopsis to your class for the first time –...Vectors at Addgene Find Simple Protocols for Introductory Lab Work MoClo Plants Parts Kit ...

...interests and areas of expertise. Perhaps the strongest aspect of science communication, then, is its ...address this obstacle in its many forms. A short stroll from my office to the tearoom would often illustrate...Cognitive LoadSummer SciComm Series: Modes of CommunicationIntro to Our Summer SciComm Series!...

...problems. For instance, it’s much more difficult to introduce a gene of interest into primary cells, so most...both purified CPP-tagged protein and an untagged control could enter cultured cells and activate expression...reporters Previous Protein Delivery Techniques: Introduction of macromolecules into cultured mammalian cells...

...reproduce someone else’s result. Then, they hit trouble. The experiment won’t work. Even if it does, they...to share with other researchers to help them troubleshoot and understand. All of this communication and... of incentives. Right now, researchers are not strongly rewarded for being transparent in their communication...