...take to achieve a clean NTC: Have a dedicated bench with a dedicated set of pipettes for ddPCR set-up. ...absolute quantification of viable lactic acid bacteria in faecal samples." Journal of microbiological methods... we have used quantitative PCR to evaluate the physical titer of our preps. We often repeat this assay...factor of 2. This means that if you set up two identical assays with the same sample, you could end up ...would be valid. This is why the AAV reference is critical. It allows you to determine if your titers are.... Each droplet contains the ingredients for amplification of the target DNA. This partitioning reduces...detection of the product. As an added benefit, replicates are built into the technology. One well (containing...

...many CRISPR applications, and they’ve also had success in clinical gene therapy applications. Read on to... but this research is still in the pre-clinical/early clinical stages. Researchers are also working to...to learn more about the current (and future) applications of lentiviral vectors! Lentiviral vector history...original system, the possibility of creating a replication-competent virus via recombination between the...of recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through...barcoding libraries have been used for complex applications like lineage tracing in heterogeneous tumors...2015: lentiCRISPR v2, used both for targeted modification and genome-wide screening.   Lentiviral vectors...

...terms of biosafety? A8: According to the American Biological Safety Association, "the two major risks ...lentiviral plasmids created for a variety of applications including cDNA expression, shRNA-mediated knockdown... looking for some additional information or clarification on these vectors. Read on to find the answers...scientists often refer to as “retroviruses” are technically gamma-retroviruses - another, separate member...differences may seem subtle they result in a key physiological difference between lenti- and retroviruses. ...transfer vectors for transient transfections? A2: Technically, yes. Will it actually work? That depends. The...when using a plasmid backbone that was designed specifically for transient expression (find some useful backbones...

...this approach has been adapted for the identification of physical materials that we use in our labs, including...RRIDs and the Resource Identification Initiative Compact identifiers for biomedical data How Addgene works...about which reagents were used? Unambiguous identification of a reagent is crucial for reproducibility...to identify and reference documents, files, or physical reagents. What are persistent identifiers? Perhaps...persistent and exist online indefinitely. PIDs historically referred to objects that exist online, but this...item. Given a specific Addgene ID # listed in a publication, you can quickly pull up the associated plasmid...plasmid page also helps researchers find other publications that have used the same plasmid through the ...

... a bacterial origin of replication (ORI) will not allow for plasmid replication in mammalian cells, for...Therefore, it is not always mission critical to make sure the plasmid replicates in the cell, as many of these... their bacterial counterparts, including how replication occurs and whether selection is necessary for... way. Selection methods, described below, are typically employed when creating stable cell lines and are...the nuclear genome, in the absence of plasmid replication, the foreign DNA will be degraded or diluted ...expressed in trans within the cell for effective replication. Cell lines expressing the Epstein–Barr virus...antigen (293E or 293T cells), allow for episomal amplification of plasmids containing the viral EBV or SV40...

...the amount of a given factor within a biological system. Basically, biosensors provide quantifiable readouts...biosensor?! You can do more than just ‘sense’ biological compounds and reactions; you can quantify them...of the biosensors used in research are very sophisticated, but at their core they can all be boiled down...pretty simply – they are systems that measure biological phenomena by generating an output which is directly...readouts for levels of different chemicals, hormones, proteins, etc., that you are interested in. The most...we will refer to as the analyte, needs to be specifically recognized by another factor, the receptor. ...fluorescence, pH, etc.). The transducer will generate a numerical value which represents the amount of analyte present...

...approach that has been the historical standard. Specifically, in our recent publications, we used the cancer ...result, how most of these “variants of unknown significance” (sometimes called “VUS”) function is unknown... if the majority of information on each cancer clinical sequencing report includes rare variants that ... variants across a large number of genes as systematically as possible. In developing these assays, we...tumorigenesis assays, drug epistasis assays, and morphological profiling can be used as generalizable strategies... are described in this paper Systematic morphological profiling of human gene and allele function ...pathway reference collection is described in this publication We also wrote an extensive review on the exciting...

...intended application - Remember, just because your antibody performs as expected in one application does ...why validation is so important. Antibodies are a critical component of the researcher toolkit due to their...major culprits in the reproducibility crisis of biomedical research (Baker, 2015; Bradbury & Plükthorn, ...strategies for antibody validation in different applications. Here, I want to highlight the general suggestions...intentionally generalizable and not specific to certain applications or antibody targets. That said, not every strategy...will be suitable for every antibody or every application. The goal is to show that your antibody has been...strategy to use will depend on your intended application and available resources.     Figure 1...

...their visions for a successful and sustainable biomedical research sector at our first Future of Research...discussed, many related to the structure of the biomedical workforce. Quotes taken directly from participants...of Medicine and the University of Massachusetts Medical School. Graduate education programs are also beginning...internship to be taken in an industry, regulatory, or clinical environment. It remains contested, however, whether...shape and inform the overall structure of the biomedical research sector, and what a tenable workforce...The panel discussion on Friday morning will be dedicated to Academic data and the Labor Market. On this...Shapes Science’ and contributing to the NIH’s Biomedical Research Workforce Working Group. Along with ...

...conflict is not always what was communicated but how it was communicated. The next step is to come up with...of lapsed communication. Making some assumptions about how something should be communicated is necessary...set of transferable skills, coupled with your technical skills, can open new career opportunities for ...conflict can also be good. In science, a reviewer’s critical comments on your manuscript is a form of (hopefully...scientist. And some amount of conflict is actually critical in the business world. Companies and organizations...traced back to one of a few trouble spots:  Communication (or lack thereof): Information sharing gets ...organization gets, and creating new avenues of communication can create as many problems as it can solve....

...Democrats more often than Republicans, the March is not about Democrats or Republicans (16). This is a Pro-...general (1). Science and politics? Science is an apolitical process for seeking knowledge. The process begins... again. No part of this process is inherently political so why is there a march in Washington, D.C., the...States government? Politics and science are intrinsically linked: governments exist in reality and science...as evidence of climate change, the safety of genetically modified organisms (GMOs), the absence of a link... grant in the last five years as polled in the American Association for the Advancement of Science) believe... it.  3) Linking scientists to one particular political party while alienating the other While scientists...

...well as protease sites to facilitate protein purification. We hope this kit will allow you to quickly ...for a variety of synthetic biology or protein purification studies in E. coli. 800 and 1900 Series If you...polycistronic E. coli expression constructs. Some applications of plasmids from this kit include the production...: Čelešnik, Helena, et al. "Biosafety of biotechnologically important microalgae: intrinsic suicide switch... Rogers, Jameson K., and George M. Church. "Genetically encoded sensors enable real-time observation ...and Charles E. Melançon III. "Detection and Quantification of Ribosome Inhibition by Aminoglycoside Antibiotics... Ribosome-Controlled Fluorescent Reporter."ACS chemical biology 11.1 (2015): 31-37. pCri system The Gomis-Ruth...

... its nuclease activity, catalytically dead Cas9 (dCas9) has many applications, the most well-known being...characterizing the 4D nucleome. Future modifications and applications Ma et al. developed this technique ...plasmids from this publication at Addgene. A protein-tagging system for signal amplification in gene expression...technique published every week! One of the newest applications is a colorful system that uses fluorescently... The technique successfully resolved loci with physical map distances of 75 and 2 Mbp, with the calculated...distances correlating with the previously established physical map. In comparing pairs of targets ~2 Mbp apart... et al. anticipate that the technique will be applicable to studies of cell cycle progression, epigenetics...

...your experiment (all well validated for your application, of course!) When the stars are aligned and the...factors that affect isotype choice. Target and application should be considered The first question you may... a strong signal and to work in a variety of applications. The desired specificity of your secondary antibody...affect my experiment?” Well, it depends on the application and target. If you are running a basic western...target is expressed at very low levels or your application requires a high degree of affinity, then species...considered when choosing an antibody. In some applications probing species-on-species, i.e. using a mouse...choosing antibodies   Complexity is a key factor Biological systems tend to be complex with a variety of ...

...show all publications by an author, total number of citations, and the number of publications that heavily... staying on top of the ever expanding body of biomedical literature! PubMed PubMed is the classic search...read the abstract or full text of the paper, publication date, a relevance score, and type of article....that term. You can also set up alerts for new publications from a particular author. Saving papers Papers...interest.  Best uses Accessing the vast body of biomedical research papers. Twitter Scientists often use... work, so it’s also a good place to look for publications. Bonus: papers have to be summarized in 280 ...organizations, and universities often tweet about publications in the fields of research they support. Monitoring...

...et. al.). Physical versus functional titer Viral titers usually come in two flavors, physical or functional...the target cell’s genome. Physical titer The two most common assays for physical titer are direct p24 measurements...functional. Physical titers measure the amount of viral particles in a sample and are usually based on the...produced can actually infect cells. These assays typically involve infecting a target cell line with your...particle. Consequently, titers based off of p24 quantification tend to be overestimates since they can include...quantify the amount of PCR product that comes from amplification of the original plasmid which can over-inflate...methods tend to be more accurate than measuring physical titer, there is a risk that they may underestimate...

... offers a significant improvement and simplification of the original technique, specifically in terms ...interaction with limited biological materials. Problems with Chip-seq ChIP-seq typically requires a large amount...material for a ChIP-seq experiment can be a major technical hurdle.   ChIP-seq requires  cross-linking during...interactions for subsequent sheering of DNA by sonification. Though it helps capture weaker or more transient...Washington. CUT&RUN doesn’t require fixation and significantly decreases background noise, reducing the read...development using the Drosophila wing as a model. A typical ChIP-Seq experiment requires around 1,000 wings...dissections. With CUT&RUN, we have had success with significantly fewer wings, such that enough tissue for an ...

...improving expression of nutritionally and therapeutically significant proteins. In this project she aims to...pests. By developing biological agents such as insecticidal proteins, the use of chemical pesticides may be...scientists work on projects spanning from the identification of plant compounds and insecticidal proteins...get tested for their potential agricultural and medical use. Find More Optogenetic Tools Like pDawn  Making...bacteria. Learn more about the different CRISPR applications in plants.     Transgenic yeast as source of... yeast capable of desaturating fatty acids. Identification of Insecticidal Proteins Linh Bao Ton (M. Sc...one of the reaction’s metabolites. One useful genetically encoded H2O2 probe is pQE-30-HyPer3 (2) which...

...techniques in the biological sciences and constitute foundational elements of biomedical research. Traditional...Restriction Enzymes + - PCR Amplification + + Digestion + + Gel Purification + - Ligation + - Transformation...Molecular cloning is the isolation, insertion and amplification of a recombinant DNA without sequence alteration... multi-step process consisting of digestion, purification, ligation, and transformation. While restriction...sequences for base pair overhangs, insert and vector purification, and, most importantly, excessive time requirements...is ligation independent, it does not require purification of insert or vector products, nor does it require...Digestion products are then transformed directly into chemically competent E. coli and transformants selected ...

...Attempts to remove excess bone surgically cause the body to “heal” the surgical site with even more bone that...therapeutic biologic design (ADCs) along with pre-clinical analytical development. His areas of expertise include...protein engineer currently working at Sanofi Pharmaceuticals in Cambridge, MA. When I joined Sanofi/Genzyme...to get in shape. After three kids, I had put on typical middle age weight and “exercise” consisted of taking...named AJ Gonzales who suffers from Fibrodysplasia ossificans progressiva (AKA FOP). This terrible disease ... But, there is HOPE. FOP is a genetic disease typically caused by a single R206H point mutation on the...growth in mouse models. As this biologic moves into clinical trials, there is new HOPE for AJ. But, as we in...