...biomedical workforce. Quotes taken directly from participants (summarized in 2) touched upon many problems...better solution would be to reduce the number of entrants into PhD programs to more closely reflect the ...Labor Market. On this topic it is clear that many important questions, areas for informed discussion, debate...Among other involvements, he helped to establish grants through the Sloan foundation for postdoc-related...

...subsequent conclusions. Being vigilant about these errors is an important part of maintaining the quality...elements, and nucleotide indels or mismatches in significant plasmid elements. Which differences are considered...This post was written by Alyssa Shepard, with significant contributions from Alyssa Cecchetelli, Cary Valley...

...cells are incubated and after several days the supernatant is collected, filtered, and titered. A major ...number of harvests will all impact the titer. Importantly, optimized conditions for one transfer plasmid...generally a good place to start. If you don’t anticipate that you will need large volumes of lentivirus...producing lentivirus, it is time to titer. One important consideration when titering a lentivirus is the...

...undergraduate students (1, 2), but educators face significant challenges in designing suitable projects. In...because the S. cerevisiae deletion strains are ∆ura3 mutants and cannot grow without uracil supplementation ... Students who participate in the course show significant gains in both content knowledge and research ...PMCID: PMC4012490. 4. Hoffman, C.S., Wood, V., & Fantes, P.A. An ancient yeast for young geneticists: A...

...decision easier. 1. Müller, Sara M., et al. "Quantification of Förster resonance energy transfer by monitoring...monitoring sensitized emission in living plant cells." (2013). PubMed PMID: 24194740. PubMed Central PMCID...) or would perturb the endogenous state that you want to measure (overexpressing a gene or protein after...detect cellular processes and signaling cascades, quantify mechanical tension (a molecular “spring”) (Meng...

...could have wanted to know was just buried beneath someone's lunch. Many times, I’m pleasantly surprised...should not) answer your questions about an Abcam antibody. Have your catalog number ready. I've attempted...last digit: An answer should come soon, but not instantly. Addgene tries to get back to customers within...

... transposon methylation underlies the mantled somaclonal variant of oil palm." Nature 525.7570 (2015):...study,epigenetic changes were statistically significantly associated with reduced life-span, but no changes...controlled, the reporter consistently showed significantly different FRET levels between unmethylated and...

...management skills but a longer term and more substantial experience can be achieved by volunteering for...national issues. I applied and was successfully granted a travel award to attend the annual NPA meeting...decide to remain here.  In a time when it is so important for postdocs and graduate students to think about...blogger Juliet Moncaster! Juliet Moncaster is an Assistant Professor at the Boston University School of Medicine...

...expressed, they predominantly fluoresce one color but slowly mature until they predominantly fluoresce a ...FP timers can be used in any situation where one wants to understand the relationship between the age of...3. Verkhusha, Vladislav V., et al. "An enhanced mutant of red fluorescent protein DsRed for double labeling...

...repairing DNA double strand breaks – (HR). HR is the dominant homology-directed repair (HDR) pathway in mammalian...higher HR frequencies than quiescent cells. It is important to know that HR is often mutated in cancer cells...recognition sequence in the donor DNA, you can guarantee there won’t be re-targeting after HDR. If your...your cells. Enrich S-phase cells HR operates predominantly in S and G2 phase of the cell cycle, when a ...

... popular and useful viral vectors in the lab. Advantages of lentivirus include a large genetic capacity...Figure 2). The HIV genes that do remain are very important for viral production: Gag (structural precursor...transduced with a lentiviral vector containing an antisense sequence against the HIV-1 envelope. As of 2013...contain a deletion in the 3’ LTR that prevents aberrant activation of nearby genes. These safer vectors...

...activator (1a) or a repressor (1b). Scientists took advantage of these discoveries to create a new system for...mating two single haploid yeast fusion plasmid transformants into a diploid strain for screening), or arrayed...increase the stringency of your binding. And most importantly: Independently validate identified binding partners...ensure that your setup works. The last thing you want is to run your lovely library screen only to find...

...the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem many...resources can be found in our Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene...looking to streamline your lab management and you want the works: organizing reagent storage and inventory... plasmid features, and reference tables (i.e. antibiotic concentrations, commonly used primers). We also...

...comparable (within r2=0.96) to that of NGS data. Advantages of ICE Determine Editing Efficiency and Types...  ICE uses Sanger sequencing data to produce quantitative, NGS-quality analysis of CRISPR editing - enabling...guide design tool, guide validation tool, and guaranteed editing with the CRISPR Gene Knockout Kit. Please...featured topic page xCas9: Engineering a CRISPR Variant with Pam Flexibility RNA Editing with Cas13 and...

... allows access to a larger number of willing participants. While fibroblasts obtained from aged patients...hiPSCs than age-matched fibroblasts [3]. These advantages make PBMCs the ideal source of somatic cells ... vectors are based on the Epstein-Barr Nuclear Antigen-1 (oriP-EBNA-1) system where each vector contains...OCT3/4, SOX2, KLF4, C-MYC, LIN28 and SV40 large T antigen (SV40LT). The method had a reprogramming efficiency...

...These differences in recombinase efficiency are important to keep in mind when deciding whether to use the...common means to achieve ligand control is to fuse a mutant estrogen receptor (ER) ligand binding domain (LDB...recombinase. The high concentration of FLEx vectors guarantees that most cells are transfected with all labeling...versions can be used in these experiments. This elegant combination of Cre, FLP, and inducible systems ...

...that asbestos exposure remains the single-most important factor in developing mesothelioma. Asbestos, and...cells that can begin an immune response. This important biomarker can be used to distinguish between three...earlier, thus improving the patient’s prognosis significantly, but also to monitor those who may be at greater...be Used to Study Cancer Drug Resistance Plasmid Grant to Aid Cervical Cancer Screening in Ghana Additional...

...into AAV. NmeCas9 has the added advantage that it can be turned off by anti-CRISPR proteins. The Sontheimer...genome modification, but they did so without a substantial immune response or toxicity. More AAV-based CRISPR...Resources on the Addgene Blog Learn more about Cas9 Variants and the Expanded CRISPR Toolbox Read about CRISPR...

... as pre-made viral preps. If you can’t (or don’t want to) make viral preps in your lab, there are custom...Titering After generating your lentivirus, you’ll want to infect your cells with the virus–the right amount...conditions to optimize include incubation time, antibiotic conditions, and amount of polybrene. (psst…didn...have a large number of cells to transduce, you may want to consider spinfection. Spinfection is the process...

...at a lower rate than proteins encoded by highly abundant, charged tRNAs. For example Tuller et al. found...ribosome will move quickly along regions calling for abundant, charged tRNAs but will stall at regions calling...! I should just choose the codon with the most abundant pool of charged tRNAs in my host organism for ...normally found at low abundance in E. coli. The advantage of producing additional tRNAs is that you can ...