...decided to pack it in. I had just started month 8 of my 3 year Post-Doctoral contract at the University of Cambridge...this job specifically looked for a researcher with 3 years+ of cancer research experience, a PhD or a Master...there a move with either promotions or further education may allow you to transition into various realms...

...exchange, the government protects (and often funds [3]) scientists as well as other citizens. Thus civilization...depending on the number of people supporting it.  3) Linking scientists to one particular political party...request of this administration promises more cuts (3, 12, 13, 27). In order to achieve the breakthroughs...sister locations on April 22, 2017, scientists and non-scientists alike will march to advocate for science...already come to pass. It may only be by advocating for science education and literacy that this changes.  2...marching for. I will be marching to advocate for science education and literacy as I believe people that...communities, microbial interactions, and science education.  Disclaimer: The views presented in this article...

...tumor-infiltrating lymphocytes of that tumor. In Figure 3 you can see how I gated out my lymphocyte population...look for them on the FSC-A/SSC-A plot. Figure 3: (A) With the acquisition voltage (not shown) used...gates while some like to make looser ones (Figure 3). A more “liberal” gate in the beginning should not... you with two measurements: forward scatter (FSC) and side scatter (SSC). To acquire these measurements...exclude all duplicate events (two cells stuck together) from your single cells, as duplicates can’t be reliably...types of scattering that occur when a cell is pulsed by the laser. Within each type of scattering, you also...to keep it short though, promise! FSC and SSC: scattering To digitally extract your cells based on their...

...Chapter 3: Finding the Right Antibody for Your Experiment.” Whatever your antibody application question...resource for anyone looking to use antibody-based applications in their research.  Like all our eBooks, the...through the process. If you’re confident in your application and protocols, but can’t quite seem to find an...

...customized sequence specificities." Nature communications 3 (2012): 1147. Cooke, Amy, et al. "Targeted...the core of a Shine-Delgarno sequence found at the 3’ end of the leader sequence. Using a variety of GFP...create custom RNA binding proteins in as little as 3 days. Abil et al., 2014 show that you can fuse custom...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...and Charles E. Melançon III. "Detection and Quantification of Ribosome Inhibition by Aminoglycoside Antibiotics...

...abundant incorporation of cysteine in their proteomes (3). This led to the eventual discovery of a naturally...which can be specifically recognized by the Sep-tRNA. 3. An expression plasmid encoding the ORF to be expressed...PMID: 25514926. PubMed Central PMCID: PMC4383998. 3. Li, Tong, et al. "Cysteinyl‐tRNA formation: the last...most inclusive databases of post-translational modifications, identifies a staggering ~250,000 phosphorylation...steric and charge-based nature of the phosphoryl-modification; they are both much smaller, and carry only ...thus cannot reflect the true state of protein modification within the context of its appropriate regulation...codons (TAG), replacing them with TAA (7). This modification allowed the deletion of release factor 1, the...

...Figure 3 shows a histogram for a yes-no dot plot, like the one shown in Figure 1.  Figure 3: An example...to the right indicates a higher signal, or upregulation, and a peak to the left indicates a lower signal...anything to the right of it, marking it as GFP+. Quantification for such graphs could simply be the percentage...case for you, switching to a contour plot should indicate where the two populations diverge. The smear ...place your gate based on the dot plot would be complicated.  Figure 2: (A) a smeary dot plot showing...Figure 4 compared to the dot plot in Figure 2. Quantification using MFI makes it easier to identify and study...addgene.org Addgene's Antibody Guide rAb Affinity Purification Protocol Video rAbs in the Addgene repository...

...used saving the researcher both time and resources (3). Still, researchers generating transgenic animals..., called dideoxynucleotides (ddNTPs), lacking the 3’ hydroxyl group required for the extension of a DNA...Medicine 307.1 (1982): 30-32. PubMed PMID: 6176866. 3. Hoebeeck, Jasmien, Frank Speleman, and Jo Vandesompele...comparison to a control sample. Southern blotting applications range from confirming the results of cloning...interest by recording DNA abundance throughout the amplification process, rather than just at the end as in standard...experiments allowed researchers to identify the location of genes. With this knowledge, researchers were...was added and when. Compiling the identity and location of a given nucleotide, a researcher (or a simple...

...incorporation of a 3’-5’ exonuclease in the reaction mixture. The exonuclease removes mismatched 3’ overhangs,...-guide RNAs – but it also allows you to skip purification steps and end-repair steps with well-designed...that the region of homology doesn’t have to be located at the absolute end of a fragment. This means that...fragment sequences and the polymerase used in the amplification. This tool allows instant updates as fragments...transforma­­­tion or as a PCR template. Some of the applications of the NEBuilder HiFi DNA Assembly Mix include...the limitations associated with otherwise more complicated assembly approaches, and opens the door for new...

...-lantern emitted yellow-green light and exhibited 3-5 fold increased brightness over previous BRET probes... single-cell and whole-body imaging.” Nat Commun. 3 (2012): 1262. PMID: 23232392 PMCID: PMC3535334 Find...resulting in a fluorescent signal. Since their first publication in 2012, the Nagai laboratory has assembled a...multicolored nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion...sensitivity and faster imaging compared to previous publications. They also developed calcium, cAMP, and ATP ...furimazine. In addition to the previously described applications, the enhanced signal of eNL allows their use...characterizing Nano-lanterns and eNL for many different applications. For long-term imaging, especially where photobleaching...

...stable gRNA than when they inserted the aptamer at the 3’ end of the gRNA. Because of the success of the MS2...can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius. Triple color detection With...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without...has been widely adapted for a wide variety of applications: base editing, CRISPR activation and inhibition... (B) CRISPR-Sirius used to visualize relative locations of the subtelomeric region of the pericentromeric...al. “CRISPR-Sirius: RNA scaffolds for signal amplification in genome imaging.” Nature Methods 15 (2018)...

...Knife, Genesis 34, 1–15 (2002).PubMed PMID: 12324939. 3. S. E. McGuire, G. Roman, R. L. Davis, Gene expression...Versatile Toolkit for Gene Expression in Drosophila, 3, 1–10 (2015). PubMed PMID: 21356876. 5. J. Chow, Z...as overexpression of each gene individually. Applications of the Gal4/UAS system There are a number of...lines. In the meantime, be sure to browse the publication references and web links from my first post and...

...positive for your marker of interest (Fig. 3).   Figure 3: Isotype flow plot example. The isotype ...ll need a few controls specific to flow as an application. These controls will allow you to distinguish...commercial antibody pages. Flow cytometry can be a complicated and daunting process. Controls like single color...

...chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment (4-chloro-3-brom-...insert. All colonies on this plate should be blue, indicating that your IPTG and x-gal are working as they ...of false positives: Blue-white screening only indicates the presence of AN insert, not necessarily YOUR...

...-C CRISPR-Cas3 for genome-editing applications. Molecular Cell, 84(3), 463-475.e5. https://doi.org/10.1016... technology and applications with the type I CRISPR system. Gene and Genome Editing, 3–4, 100013. https...repression, high precision edits, and more. Figure 3: Schematic of the basic CRISPR-Cas9 mechanism, representing...double-stranded breaks caused by Cas12 create short 3’ overhangs, which leads to slightly increased efficiency... your own research! Classification Structure Principles of CRISPR classification While CRISPR systems...Shakespeare for the first time. The discovery and application of CRISPR-Cas9 kicked off over a decade of frenzied...properties. This led to a need to develop a classification system for the array of CRISPR technologies...

...resulted in a roughly 6-fold increase in expression [3, Figure 2]. Note how expression increases exponentially...PMID: 18500252. PubMed Central PMCID: PMC2574934. 3. Zingg, Brian, et al. "AAV-mediated anterograde transsynaptic... Protocols for Work with Virus Watch Our AAV Purification Video ...

...some impressive videos of these interactions (Ref 3). You can find useful tips on in vivo imaging in our...Research 9.12 (2016): 895-905. PubMed PMID: 27913448. 3. Cai, En, et al. "Visualizing dynamic microvillar ...Immunosurveillance and Defense With increasingly sophisticated visualization techniques it is possible to follow...

...no addition of nucleotides to the 5’ spacers. The 3’ ends were modified slightly with either a 1-7 base...constructs, transcript levels of PTG1 and PTG2 were 3 and 31 times higher, respectively. In rice protoplasts...biological pathways;  to observe a phenotype, the modification of multiple genes is often necessary. Guide ...recognition and cleavage. This sequence flexibility indicates that gRNAs could be incorporated into these constructs...the 5’ end. PTG construct schematics. Scissors indicate cleavage sites. After confirming gRNA cleavage...) How many loci can Cas9 modify? To test the application of this system to gRNA multiplexing, Xie et al...deletion with 4-20% frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct...

... cover letter can be found on the Addgene blog.   3. Personal statement brilliance In addition to sending... Letters of recommendation wisdom  Reach out to 2-3 individuals in your professional circle to vouch for...your application a whole week before the deadline. The day of submission, review your application with... their dream.  To help students focus on the application process, and simplify access to academic resources...Scrutinize every opportunity prior to starting an application to assess your eligibility, interest, and career... detailed. Beyond the resume, a cover letter communicates your interest in a specific program, and institution...February represent peak season for internship applications. Thus these noble individuals are inundated ...

... the pH change caused by storage on dry ice (Fig. 3). However, 30 seconds was not sufficient to prevent... was a substantial reduction in infectivity after 3 weeks of exposure to pH 5.1 (Potter et al., 2014)....improve our shipment processes. After reading a publication describing how short term storage on dry ice ...medium, which is buffered and contains the pH indicator phenol red. We froze samples in a -80°C freezer...closed dry ice storage container for 48 hours (to replicate how our viruses are shipped), where they would...exposed to CO2/dry ice. All samples were tested in duplicate. What we found Not all tubes (or rather, not ... the samples were at the control pH of 7.4 as indicated by color (data not shown). What we did next: Venting...