...multiple DNA fragments into many vectors at once in the same tube. You can clone up to 4 DNA fragments, in a .... Universality All types of DNA fragments may be cloned: PCR fragments, cDNA or Genomic DNA and is available...reaction creates an expression clone with all of the components necessary for gene expression.     The LR...important to include the proper protein expression elements (ribosome recognition sequences, start codon, ...transfect the cells that you want to use for your experiments and verify that your construct is functional....recombination; ccdB gene for negative selection; elements to express the gene of interest in the appropriate...

...directed evolution approach with some notable improvements. Directed evolution techniques mimic natural...utility of the system for use with low affinity variants of TEV. The new system has been demonstrated to...the evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription...fluorescent tools for labeling structures and compartments in mammalian cells By Angela Abitua Genetically...are often used to label  specific structures, compartments, or specific localization of biomolecules within...plasmids at Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases... allow for sensitive and high spatiotemporal measurements of norepinephrine. These norepinephrine sensors...

...similar situations to themselves, allowed the participants to feel more comfortable at a large meeting....They really liked us.  The testimonials from participants in Binning Singletons at ASM Microbe in 2019...people to talk to and attend sessions with.” These sentiments were echoed in the other testimonies we gathered...people to do in the pilot. We also found that all participants would benefit from more structure with regard...assistance than others, but we did have some participants feel like their mentors weren’t as invested ...expectations have been established for future participants regarding the activities and time mentors spend...Binningsingletons.com will be the hub for all our events, tips, and more. We are partnering with ASM and...

...major benefit when recruiting patients, specifically children and patients with skin disorders for research...larger number of willing participants. While fibroblasts obtained from aged patients are reprogrammed less...iPSCs: Blood is easily and routinely drawn from patients in contrast to the process of collecting fibroblasts...correlation has been found between the age of the patients from whom the blood is obtained and the subsequent...reprogramming factors behave as separate extrachromosomal elements in the nucleus of the target cell. The method ...-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science, 2003....

...such as adjuncts) and graduate and undergraduate students are exempted from the law. However postdocs, staff... effects on higher ed had been seized upon by opponents of the ruling. However when the ruling was issued...Overtime Rules, made clear that institutional arguments were not as convincing as the recommendations ... therefore the numbers of postdocs at different points on this salary range. We have been able to gather...the institutions that are meeting the salary requirements of the FLSA. In the University of California...get rid of postdocs; that current postdoctoral appointments may shorten in length; that postdoc salaries...accept these recommendations, and make gradual adjustments towards this new level. Indeed some institutions...

...important binding events take place during the course of infection. These binding events can be impaired...the most out of your lentiviral transduction experiments. Love your cells and they'll love you back ... problem that often plagues labs and hinders experiments is mycoplasma contamination. Mycoplasma contamination...cloudy. Mycoplasma compete with host cells for nutrients and can alter expression of receptors, ion channels...collect and pool the harvests from multiple time points while others opt for a single harvest after transfecting...of infection (MOI) for predicting gene transfer events.” Genetic Vaccines and Therapy 2.1 (2004) Pubmed...Titering Your Lentivirus Learn All About Viral Vector Elements Learn How to Do Genome-Wide Screens with CRISPR...

...some types of research, raw data requires huge amounts of storage. For example, mouse behavioral geneticists...different occasions via video, generating huge amounts of data. When published, this data is usually condensed... of data is essential for efficient science. Improvements in our discipline for annotation and archiving...not finalized its definition of necessary data requirements and likely will not do so (calling it a “sisyphean...another issue of proprietary data formats. Many instruments store raw data in formats that require specific... and Dataverse. Just as Addgene helps with DNA reagents, these resources can offload the burden of data...rely on many other labs to teach us how to do experiments and understand their data. Likewise, we want ...

...chat for Early Career Researchers.  Many of the participants had certainly realized the importance of seeking...What makes a good Mentor?” Some of the Tweeted comments: “A good mentor has your goals/needs/situation...most of them Is not threatened by the Mentee’s talents or ambitions Encourages the Mentee to take risks...thoughtful treatise on the kind of advisor/mentor she wants to be for her new lab. If only every science trainee...their career. “I want to be a PI that holds my students accountable for their actions and pushes them ...without being taken advantage of, and to respect my students as developing colleagues even as I reinforce the...the necessary power dynamic between advisor and students.” Stay tuned for the answers to the other six ...

...Their small numbers do not, however, make these patients suffer any less acutely.  The Rare Genomics Institute...those who suffer from rare diseases by connecting patients to important resources and promoting research ...disease. This year, Addgene will be providing plasmid grants worth $2,000 each to two labs through the BeHEARD...accelerate rare disease research and provide hope to patients suffering from these devastating afflictions. ...motor neuron function in spinal muscular atrophy patients. Noushin Nabavi - Peritoneal Mesothelioma  Noushin...mesothelioma, there are currently no targeted treatments for peritoneal mesothelioma. Dr. Nabavi and colleagues... lines and assess their tumorigenesis. These experiments will serve to validate these candidate genes ...

...As previous blogs have noted, plants are an important foundation for life on Earth. Selective breeding...breeding methods have shaped the plants that we grow and eat, and genetic engineering will continue to improve...transient and stable gene expression applications in plants. This smaller, easily customizable vector functions...biolistic methods, making it useful for engineering plants that are not susceptible to Agrobacterium infection...select both transformed Agrobacterium and infected plants. Further opportunities with pSiM24 While these ..., three MCS regions permit multiple regulatory elements to be inserted, further customizing gene expression...as well as stable expression of foreign genes in plants." PLoS one 9.6 (2014): e98988. Pubmed PMID: 24897541...

...first in a series that will feature advice for students, postdocs, and young Principal Investigators (...alerts for new papers with keywords or a Table of Contents email (eTOC) alert for every new issue of your...careers outside of academia Recently, graduate students and postdocs have become more aware of careers...all the real life skills that employers love PhD students to have: working to deadlines, writing up clear...their personal goals to their career choices.” She points out that a PhD teaches analytical and critical ...outside of academia. She has had previous graduate students and postdocs go on to work in patent law, business... Your Lab,” “Fun in the Lab!" and more.   Acknowledgments Thank you again to Connie Cepko, Tom Ellis...

...integrase mutants in the early 1990s (Leavitt et al., 1996), and since then, numerous variants have been...These trials unfortunately resulted in several patients developing T-cell acute lymphoblastic leukemia...titering IDLVs include detecting vector particle components or performing quantitative PCR titration of vector...research, we wish you the best of luck with any experiments involving IDLVs. And remember: You can never ...Consequences on Host Gene Expression. Viruses: Essential Agents of Life, 147–175. https://doi.org/10.1007/978-94...leukemogenesis following gene therapy of SCID-X1 patients. The Journal of Clinical Investigation, 118(9)... Human immunodeficiency virus type 1 integrase mutants retain in vitro integrase activity yet fail to ...

...DNA-based reagents are named for soup ingredients! From pSOUP to mCherry, satisfying ingredients reagents...Gram-positive bacteria (Kellenberger et al., 2013). Other variants of Spinach include Baby Spinach, a miniaturized...short non-coding RNAs (Autour et al., 2018). Three variants of Mango, coined Mango II, III, and IV, were then...high binding affinity to TO1-biotin. Two of these variants exhibit increased brightness compared to Mango...useful for both live and fixed cell imaging. New ingredients in the aptamer soup In addition to these established...

... and quality for HIV-1 patients. However, ART is not an HIV-1 cure. In patients who stop ART, viral levels... latently infected cells. Although ART-treated patients lack plasma HIV-1, they are at increased risk ... isolated CD4+ T-cells from two HIV-1 infected patients. Although this is an exciting finding, it’s important...important to note that these HIV-1 patients were ART-naive, so this experiment models the active HIV-1... ART-induced viral control found in most HIV-1 patients. Obstacles for CRISPR HIV-1 therapy Both of these...CRISPR gRNAs require DNA sequence-specific binding. Patients’ HIV-1 genomes will need to be sequenced to determine...discouraging, it’s important to note that these experiments were done in cell culture, and the cells infected...

... Designs Consulting (EDC), we instead have our clients focus on their experimental needs and consider ...accomplish your science, and can include: equipment, reagents, animals, glassware, and even people. When developing...even includes clocks, timers, and phones.  As for reagents and consumables (such as pipet tips, glassware...to need a tech because you’ll be doing mouse experiments, start defining keywords you’ll want to find ...their goals, rather than focusing on long term investments. For example, purchasing “used” equipment can...resource budget, and know how to decide which components are the most critical to your research, negotiations...limitations. Everything is negotiable. Try asking for discounts, collaborations, access to internal award applications...

...some extra guidance to understand the specific components that make up a plasmid and why each is important...spend more time developing cleverly designed experiments and innovative solutions necessary for advancing...naturally in archaea and eukaryotes such as yeast and plants. In nature, plasmids provide one or more functional....   Figure 1: Map of a plasmid with its elements described below.   Vector Element Description...as they grow, scientists can easily make large amounts of plasmid to manipulate and use in later work....plasmids can be used to do include: Produce large amounts of a protein so that scientists can purify and ...Plasmids 101 series, please let us know in the comments. Note:Marcy Patrick contributed to the writing...

...awareness for rare disease and helps patients find the correct treatments, if available. As part of the team...play with my kids like any other youngster. His parents, Kristi and Rico, had to keep him from running ...triggers appropriate bone regeneration in healthy patients. However, the R206H mutation makes ACVR1 responsive... with inappropriate activin-ACVR1 interactions prevents FOP bone growth in mouse models. As this biologic... But, as we in the scientific community know, treatments that work well in animal models do not always... Disease Researchers Check Out Potentiall HIV Treatments with CRIPSR Resources on Addgene.org Find ACVR1...

...each level, as each level has different safety requirements. BSL-1 is designated for those working with ...

...to address the comments to “you” rather than “the authors.” By addressing the comments to “you” I can ... figures). After finishing my section-specific comments, I re-read what I wrote about the paper after ...in the first (and clunky) version of my general comments and adjust them if my opinion changed after this...that there is another human who is reading your comments. Receiving a peer-review is already a humbling...young scientists that are reading your peer-review comments will learn from them and propagate this culture...ratio in Arabidopsis, (2) expression patterns in plants with enhanced sodium accumulation in their roots...

...oligomerization can lead to misleading artefacts in your experiments. Read this post from Erik Snapp, Associate Professor...oligomerization can be and how to avoid it in your experiments. Better Dyeing Through Chemistry & Small Molecule...fluorophores can still be used in a wide variety of experiments. Read more to find chemical fluorophores for ...and/or engineer “temporally ordered biochemical events” such as those that occur during development. Read...study basic virus biology while leaving virulent components behind. Minigenomes are stripped down viral genomes...fuel sources.” In her work, Dr. Polka made R-body mutants that respond to different pH’s and some that are...libraries of CRISPR gRNAs to use for screening experiments. Get the full details on how to make these libraries...