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...the learning of many facts but the training of the mind to think something that cannot be learned from textbooks... (Brian, 1996). But is it possible to bring the kinds of transformative educational experiences afforded...happened in the lab, and he wanted to provide the same kinds of opportunities for his students. But he also found...genuine interest to other researchers in their field. Indeed, sharing results with the broader scientific community...interact with citrate synthase and presented their findings at the American Society for Biochemistry and Molecular...research, they gain confidence and learn to work independently, which can lead to greater success in meeting...working, hypotheses being rejected, etc.), while reminding participants of the value of this approach and...

...our Fluorescent Protein Collection Find Plasmid Kits at Addgene Find Plasmid Cloning Protocols References...promoters, enhancers, polyadenylation signals, inducible gene-expression sequences, and more. By combining...constructs to visualize cell cycle dynamics, titrate inducible transgene expression, or a variety of other clever...label subcellular structures in live cells. Four individual expression constructs were created, each containing...Repeatable, directional cloning   The principle behind MXS-chaining is repeatable, restriction-enzyme ...cloning. The modules (Figure 2) are contained in individual plasmids and are each flanked by the same multiple...

...protocols for plasmid cloning Find our molecular biology reference here Find molecular biology protocols...bacterial recombination pathway, referred to as recA-independent recombination, joins together pieces of linear...sequences! Primers consist of two regions: the 3’ end binds template DNA for PCR amplification, while modifications...encoded at the 5’ end. First design the template binding region, with a melting temperature (Tm) of around... as epitope tags, design primers with template binding regions to amplify away from the insertion site... To remove part of a plasmid, order primers that bind at either side of the undesired portion, designed...

...expression in mammals Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all...horribly wrong. It brought the whole field to a grinding stop and only now, 2 decades later, is it finally...autosomal recessive, severe, childhood-onset form of blindness name Leber’s Congenital Amaurosis (LCA). Shortly...disease that results in combined deafness and blindness) is caused by defects in the gene MYO7a, at ~7...further reading 1. FDA approves gene therapy for blindness 2. Gene Therapy emerges from disgrace to be the...restores vision in a canine model of childhood blindness. Nature Genetics 28, 92-95. PubMed PMID: 11326284...

...on Addgene.org Find CRISPR Plasmids for Your Research Read Our CRISPR Guide Pages Find CRISPR gRNA Design...bacteria causing an infection, but also alter an individual’s native, beneficial flora. Currently, there is...-homologous end joining machinery, so a CRISPR-induced double-stranded break (DSB) in the genome is lethal...be eliminated from the bacterium, which may also induce cell death. CRISPR also enables increased specificity...sequence-specific, antimicrobial, plasmid-based, CRISPR systems (Find the Citorik Plasmids here). In both cases, they ...PMID 252409288. PubMed Central: PMCID PMC4˚237163. Find plasmids from this paper at Addgene. 2. Bikard,...

...priorities to attend to. Being mindful looks like: “When you have time, would you mind helping me with….” vs. ...become proficient in lab techniques, which will hinder your progress. You will also probably have a lot...there enough information there for you to be independent performing the technique you trained to do? Walk...clicking’ for them. It’s often easiest to train individuals who are polite, appreciative, and clear about... be done by the researchers themselves.  Be mindful that your mentor will still have their own deadlines...Have a Mentor, What Now? Will You Be My Mentor? Finding and Asking for Mentoring Support   ...

...organisms - organisms have codon usage bias. You can find many codon tables showing which codons encode which...preferences and what can we do about them? Read on to find out!   Why do organisms have different codon usage...instance, it may not express at all in E. coli (you can find a variety of databases showing various organisms...codon choice may interrupt transcription factor binding sites) As you might imagine, it’s not easy for...GenScript host online codon optimization tools. Keep in mind that, just because you optimize a gene with one ...et al. "Selective charging of tRNA isoacceptors induced by amino‐acid starvation." EMBO reports 6.2 (2005...PMCID: PMC552224. 4. Gustafsson, Claes, Sridhar Govindarajan, and Jeremy Minshull. "Codon bias and heterologous...

...Learn about Addgene's viral service Find viral protocols and videos Find AAVs for your research ...Locomotion is a highly complex process on the level of individual neurons and in terms of their interactions with...like stress levels, but researchers have to be mindful of motivation overlap. The locus coeruleus (LC)...2015) used optogenetics to support these existing findings by controlling neurons in the mPFC with light....depression, yet there have also been contradictory findings (Groves, 2007). A study from 2014 (Bahi et al....Addgene blog Read all of our viral vectors blog posts Find tips for getting started with viral vectors for ...

...clients. In this realm, there are a few different kinds of challenges that consultants get called upon to...healthcare consultant, I have worked on most of these kinds of projects. Almost all of my projects, have involved...commercial future, and yet are incredibly difficult to find solutions to. There are many reasons why client ...particular therapy. With your scientific knowledge behind you, you’ll have the opportunity to learn the business...those conversations, reading online healthcare industry publications like FiercePharma, FierceBiotech,...about the trends and workings of the healthcare industry. Finally, on-campus resources like career offices... both provide up-to-date information about the industry, recruiting processes, and can connect you to ...

...your own, but if you ever find yourself in need of a few ideas, I hope you’ll find this post useful. After...this, I always make sure I have explicit goals in mind for the endpoints of a task or project (with metrics...hand write to-do lists at the start of each day to remind myself of the specific activities that I need to... better when I write them down. Some people also find it useful to write out their to-do lists at the ...handwriting, is copied below): Read new Zhang Lab Paper Find plasmids similar to those in the paper Ask Mary ...everything you need to do by 6 pm. Hopefully you’ll find this post useful for making your days, experiments...

...some creative approaches to lab sustainability, and find ways to further reduce our consumption beyond recycling...energy, and water use. ACT labels are obtained by independent audit, and companies need to pay to obtain them...grades of plastic lumber and distribute it to industrial and residential markets. Rethinking our approach...practices to meet higher environmental standards. Find some easy action items on our other article in this...initiatives for the lab Subscribe to the Addgene blog Find lab tips for your research Resources on Addgene.org...other scientists save resources by sharing plasmids Find plasmids at Addgene ...

...to Addgene to find the plasmid or viral vector for your next experiment, that you can find what you need...finally share with you what we’ve been working on behind the scenes here at Addgene.  Addgene's new homepage... content was either small, cluttered, or hard to find. This made the page look not only confusing, but...browse functionality. We wanted users to be able to find plasmids and viral vector tools more easily. In ...what Addgene has to offer, and to help scientists find what they need - whether it's their first or 500th...

...Addgene.org Find CRISPR Plasmids for use in Mammals Check out Our CRISPR Guide Pages Find Plasmids for...have been accumulated over the years using them. Indeed, the future of mouse work is bright as it is now...if we could just go in and quickly manipulate individual mouse genes and study their function in health...this experiment showed a mutation! I looked out my window and I saw the Dorr Mountain with the rest of the...-homologous end joining (NHEJ), this will leave behind a “scar”, with nucleotides missing or added at ... when generating knockout models with CRISPR and find that many, if not all, the mice carry a frameshift...

...   NEBCutter2: For finding restriction enzyme cut sites Webcutter 2.0: For finding restriction enzyme ...enzyme cut sites Open reading frame finder  Addgene vector database: Find sequences for plasmids assembled ...Reagent and Data Repositories These links can help you find experimental and/or validation data on molecular...Addgene.org: Read our Molecular Biology Reference Find molecular biology protocols Browse our protocol ...

...it easier for you to find and use the plasmids you need. Here’s what you’ll find in this article: CRISPR-based... generation, controllable gene knockouts, and inducible gene inactivation or activation in an organoid...Organoid EasyTag system, showing the workflow for inducible CRISPR inhibition (CRISPRi). A) Lentiviral vector... to the petunia SL enzymatic receptor DAD2. SLs induce a conformational change of the receptor which modulates...plasmids are always being added to the repository. To find all of the CRISPR plasmids available from Addgene...

...Both D10A and H840A are potent editors, but their indel profiles vary. In the PAM-out configuration, D10A...nickases for HDR is targeting range: using an individual gRNA with WT Cas9, repair levels decrease rapidly...rapidly 10 bp from the cut site. So if you can’t find a good gRNA that cuts close to your insertion site...throughout the entire region between the nicks? To find out, Schubert and Yan designed a PAM-out nickase... to using WT Cas9 with the left gRNA across the window. In a subsequent experiment, they were also able...Resources on Addgene.org Check out CRISPR Topic Page Find CRISPR Nickase Plasmids Browse the CRISPR Guide ...

...activated with light.  Find plasmids from the Isacoff lab! They then tested what kind of swimming behavior... ready-to-use viral service Find optogenetics AAVs for your research Find all AAVs from the viral service...even be expressed simultaneously and still be individually activated with different lights (Baier et al...PMC2770190.  Additional resources on the Addgene blog Find all our optogenetics blog posts Read blog posts ...

...initiation of a vast number of signaling cascades, inducing different cellular responses. One of the biggest...complexes that are activated in response to a ligand binding a GPCR of interest. How does TRUPATH work? The ...bound to the intracellular surface of the GPCR. Binding of a ligand to the extracellular surface of the...bioluminescence resonance energy transfer 2, or BRET2. With binding of the drug, the alpha and beta subunits dissociate...Browse all of our fluorescent protein blog posts Find other fluorescent biosensors Resources on Addgene.org...Browse the catalog of fluorescent protein plasmids Find biosensors for your experiments ...

...of vectors for inducible expression (pSBtet derivatives). The constitutive and inducible expression vectors...transposase enzyme is expressed (red stars) and binds the inverted repeats (green); and an endonuclease...cuts the DNA.3.) The released transposon can now bind a strand of DNA with a TA dinucleotide (there are...Jan;257(1):181-90. doi: 10.1111/imr.12137. PubMed. Find the Plasmids at Addgene Kowarz Lab plasmids Izsvak... inducible vectors...

...and published by academic scientists. To help you find the vector that fits your experiments, I've described...close as possible to the basal expression level, induce expression at an opportune time or go all out with...In most cases bacterial expression vectors are inducible (e.g. pBAD LIC cloning vector (8A)) and have an...be released from the resin and used for various binding or folding assays. Yeast vectors There are several...integration into the fly genome and are under the Gal4 inducible UAS promoter. Other vectors can be expressed in...the comments below and an Addgenie will help you find what you need.   More Plasmid Resources from Addgene...