...raise salaries prior to the injunction are still honoring these plans. We are now collecting data on the...who had a contract - negotiated by their union - securing higher salaries. This decision will carry over...geographical regions When we originally started gathering this information, many assumed that those living...Read Gary's Previous Article: Changing Labor Laws Bring Increased Postdoc Wages Read about the Advocating...

...large scale cell mobility are essential processes during embryonic development. These are required for a...development of cell sorters together with the engineering of fluorescently labelled antibodies made it ...)? How do blood monocytes patrol blood vessels during immune responses (Auffray et al. 2007, Finsterbusch...relatively little tissue absorption, weak tissue scattering and small amounts of tissue auto-fluorescence...understanding the key roles of immune cells in triggering an immune response. New fluorescent proteins ...Dynamics of neutrophil migration in lymph nodes during infection." Immunity 29.3 (2008): 487-496. PubMed...Jackson G., et al. "Macrophage and T cell dynamics during the development and disintegration of mycobacterial...

...Addgene Blog Mentoring for Scientists Series. I have been thinking a lot about Mentoring for over 10 years...group mentoring format could work wonders.  I have now seen the formation of hundreds of mentoring groups...also serve as a great first-time mentoring experience. They make mentoring less mysterious and train the ...useful for future mentoring relationships.   If you would like to start a group mentoring program at your...interested peers and form your own peer mentoring circle. In a Peer Mentoring Group all participants act as both...and what you want to cover during the year. Suggest that each person bring 3 topics and write them on ...involved in Peer Mentoring relationships? Comment below with ideas from your own mentoring group or with ...

...to learn about her mentoring style. Top 7 questions scientists ask me about mentoring In this blog series...the other six mentoring questions in future posts. If you have a question about mentoring that you want... enough?  For many years I’ve been organizing mentoring programs for scientists and doing training to ...fulfilling, productive relationships. I will be sharing what I’ve learned along the way in this ongoing... to the top questions scientists ask me about mentoring. Here they are: What makes a good Mentor? How...? What is this I hear about “group” or “peer” mentoring? How do I ask someone to be my Mentor? I have ...certainly realized the importance of seeking out mentoring as an important aspect to their ongoing success...

...bacterial CRISPR engineering One common way of accomplishing bacterial genome engineering is done with recombineering...combines CRISPR and recombineering-based MAGE (Multiplex Automated Genome Engineering) technology (Ronda...combines recombineering with CRISPR to create a system for scarless, iterative genome engineering (Jiang...promoters in Pseudomonas syringae and Klebsiella oxytoca. Microbiome engineering Scientists have begun to...discovered in bacteria, most CRISPR-based genome engineering has taken place in other organisms. In many bacteria...quirks change the way CRISPR-mediated genome engineering functions in bacteria, but have no fear - plasmids... recombineering, a technique that utilizes phage recombination machinery to promote homologous recombination...

...KTRs are not well suited for monitoring kinase activity that occurs during nuclear envelope breakdown (...important for regulating signal transduction and measuring kinase activity at the single-cell level can aid...signaling activity and cell phenotype. One method for monitoring live single-cell kinase activity is FRET, but...Advantages of KTRs When compared to FRET reporters for measuring kinase activity, KTRs have two key advantages....

...multiplex genome engineering. Image from Sakuma at al. (2015). Multiplex genome engineering demonstrated...system for CRISPR/Cas9-mediated multiplex genome engineering is now available thanks to researchers at Japan...making an all-in-one vector for multiplex genome engineering.” Now, they’ve filled that gap. As Sakuma and...efficient viral vector-mediated multiplex genome engineering. The system now provides an efficient targeting...Sakuma’s protocol. Reference: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector... multiplex genome engineering...

...database and ordering platform soon to be released. For individuals interested in ordering from ABRC for...Learn how BioBuilder can bring Synthetic Biology into the classroom Engineering the Plant Genome Using ...to us at [email protected] for help setting up an ordering account. Using Arabidopsis in the classroom has...CRISPR/Cas9 pSiM24: Simplifying Plant Genetic Engineering Resources on Addgene.org Find Plant Vectors...

...platform for sharing, annotating, and storing life science protocols. Since we began sharing this protocol...Developing Lab Management Software    Organizing and sharing your updated protocols So that’s great, but what...Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene? Check out all the benefits...feature further simplifies lab work if you like to bring your phone in the lab, though, honestly, this still...non-bench work – but is there a sterile, clean way to bring your phone into the lab? Maybe having a dedicated... protocols listed on OWW, including those for preparing your own competent cells. If you’re familiar with...exchange currently boasts over 2,500 protocols covering a variety of bio and biomedical topics. Protocol...

...scientists have modified for a wide range of genome engineering techniques . Collectively, these techniques are...R epeat) technologies. Before CRISPR, genome engineering approaches like zinc finger nucleases (ZFNs) ...CRISPR rapidly became the most popular genome engineering approach. Figure 1: Overview of the basic CRISPR...first and come back to HDR . NHEJ is a naturally occurring repair pathway and is the primary repair mechanism... for DSBs. While this pathway is efficient in repairing DSB, it frequently causes small nucleotide insertions...: Double-Strand Break (Cut) Multiplex Genome Engineering Many CRISPR experiments involve editing more ...more than one gene. In these cases, delivering multiple gRNAs using a single plasmid ensures that all gRNAs...

... O 2 ) Monitoring of basal H 2 O 2 levels with peroxiredoxin-based probes Real-time monitoring of basal...81. Loren Looger Calcium NEMO calcium sensors Engineering of NEMO as calcium indicators with large dynamics...sensors, ER-targeted CEPIA1er ER-mitochondria tethering by PDZD8 regulates Ca(2+) dynamics in mammalian...sensors ZapCY1 and ZapCY2, Golgi or ER targeted Measuring steady-state and dynamic endoplasmic reticulum...alternately colored FRET sensors for simultaneous monitoring of Zn(2)(+) in multiple cellular locations. PLoS...Improved ATP FRET Sensor For Yeast Shows Heterogeneity During Nutrient Transitions. ACS Sens. 2020 Mar 27;5(3...-0. Yulong Li ATP/ADP Monitor the ATP:ADP ratio during live-cell imaging (Perceval) A genetically encoded...

...using glass cloning rings to isolate colonies of antibiotic-resistant cells; during the time required to... to grease, place and fill a dozen cloning rings, the remainder of the colonies on the plate dried out...out and died. The alternative to cloning rings, dilution cloning into 96-well plates, is extremely time...minutes- a far more rapid pace than using cloning rings. A picture of a 60 mm dish ready for picking is ...differ grossly from those isolated using cloning rings in terms of growth rates, morphology or other physiological...

... high-quality quality control measures such as titering, purity assessment, sterility testing, and viral...Request AAV Packaged on Request Overview Details Ordering Share Your Results Eligible Plasmids Additional...levels. We offer 961 in-stock viral vector preps, delivering high quality and ready availability at affordable...Addgene’s comprehensive quality control process: Titering Purity assessment Sterility testing Viral genome...chosen plasmid on its material page. Request and Ordering Process AAV production will take approximately...requests will be approved within two business days! Ordering At your convenience After approval, the next step...AAV Production 6–8 weeks Quality is our priority. During this time, we amplify DNA, produce the prep, and...

...customers. Details about our production protocols, titering methods, and quality control are described below... or through collaboration with viral vector manufacturing facilities, such as the University of Pennsylvania...are then aliquoted and stored at -80 °C. Titer Titering is either performed by Addgene or by the University...University of Pennsylvania Vector Core. In general, titering is performed by the facility that produced the...those achieved using the standard qPCR method. Titering by the University of Pennsylvania Vector Core ...Purity Purity of AAV preparations is assayed by comparing the relative stoichiometric ratios of the viral...aliquoted, frozen, and stored at -80 °C. Titer All titering is performed on lentiviral preparations that have...

...nuclease fusions Related Genome Engineering Technologies TALEN Engineering Reagents CRISPR Plasmids...Find reagents for engineering and expressing zinc finger proteins deposited by Zinc Finger Consortium...have deposited at Addgene various reagents for engineering and expressing zinc finger proteins for distribution...reagents are provided below: Zinc Finger Genome Engineering Reagents Zinc Finger Arrays for Zebrafish Genes...fingers targeting specific genes Oligomerized Pool ENgineering (OPEN) Reagents - Engineer zinc finger arrays...arrays via OPEN method Modular Assembly Engineering Reagents - Engineer zinc finger arrays via Modular Assembly...

...thaw of antibodies and suggests preparing single-use aliquots before storing at -20 °C. Each antibody catalog...production, all antibody preps are titered and undergo stringent quality control by Addgene or a trusted quality...distribution. Details about our production protocols, titering methods, and quality control are described below...Coomassie staining, the sample is separated via a denaturing SDS-PAGE gel alongside serial dilutions of a ...each order based on the total items in the order, ensuring each item ships safely and most economically. ...

...before starting your experiment. Genome Engineering Genome Engineering Overview CRISPR Cre-lox TALENs Zinc...Kit Types Fluorescent Proteins Cloning Genome Engineering Pathways & ORFs Synthetic Biology Pooled Libraries...plasmids all built with the same backbone and only differing in a small region. Click here to view all pooled... Cell Migration Consortium Center for Genome Engineering EXtracellular Plasmid RESource (EXPRESs) Consortium...

...Plamids for Oligomerized Pool ENgineering (OPEN) method has been used to engineer ZFNs that function ...used to engineer ZFNs via the Oligomerized Pool ENgineering (OPEN) method. This kit consists of 5 individually...of Kit Original Publication Oligomerized pool engineering (OPEN): an 'open-source' protocol for making ...Joung et al., PNAS 2000 ). The Oligomerized Pool ENgineering (OPEN) method has been used to engineer ZFNs ...For your Reference section: Oligomerized pool engineering (OPEN): an 'open-source' protocol for making ...

...SynBio Assembly Standards SynBio Blog Genome Engineering SynBio Depositing Labs Hal Alper J. Chris Anderson...Weiss Synthetic biology describes the rational engineering of biological systems for novel functions. Addgene... biologists around the world to facilitate the sharing of parts and information. Addgene can help you ...Regulation Browse plasmids containing both naturally occurring and synthetic regulatory elements, including promoters...Bio-Energy Institute Synberc - Synthetic Biology Engineering Research Center The PLOS Synthetic Biology Collection...

...re-purposed for site-specific eukaryotic genome engineering. CRISPR/Cas9 is an inexpensive, facile, and efficient...site. Design additional primers to analyze for scarring (small indels produced at the sgRNA cleavage site...to amplify the sgRNA target site to examine for scarring. This may be useful to characterize the non-deleted...Minimize the time between electroporation and transferring the solution into media to enhance cell viability...of Transfected Cells Prepare cells for FACS by filtering them through a 50 μm filter into a FACS tube. ...manuscript here: Bauer et al, 2014 322.2 KB Genome engineering using the CRISPR-Cas9 system. Ran FA...