...as a control. DNA was analyzed by PAGE. The basis of the Surveyor assay is that Surveyor nuclease ...edit. Using Surveyor nuclease followed by gel electrophoresis, these mismatches are indicated by the presence...Figure 4). 5. In theory you could use densitometric analysis to quantitate the intensity of the bands on your...

...plasmid vectors and/or rely on cumbersome in vitro synthesis of single guide RNAs (sgRNAs), are time-consuming...reagent preparation in the lab. Figure 3: csRNP-assisted epitope tagging is scalable to 96-well format...., Gogolok S et al. 2017. Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic...

...undesirable side products generated during chemical synthesis of these molecules. To make large amounts of pure...applications, but there are more! Engineering disease resistance through gene stacking, increasing production ...Generation of 8× Nicotiana Benthamiana and 12× Arabidopsis Mutants.” The Plant Journal, vol. 106, no. 1,...

...depends on it. Deanna McLafferty is an editorial assistant and activist. Washington, DC, USA 38.9072°N 77.0369...scientists – including global warming skeptics – insisted that more data and research funding are critical...trying turn a few exoplanets into a successful PhD thesis. We use the same atmospheric physics to understand...

...number. First you generate four unique kanamycin-resistant plasmids, each containing a different gRNA target... all but one of your gRNAs into spectinomycin-resistant plasmids pX330S-2 to pX330S-(last gRNA number).... Yin, Linlin, et al. “Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs...

...engages the prime editor protein. The prime editor consists of a Cas9 nickase fused to a reverse transcriptase... domain for optimization. They developed phage-assisted evolution strategies to evolve more effective ... J., Osborn, M. J., & Liu, D. R. (2023). Phage-assisted evolution and protein engineering yield compact...

...creation of improved fluorescent proteins through mutagenesis. Chapter 6 - The future of FRET and fluorescent...guest, Joachim Goεdhart! Joachim Goedhart is an assistant professor at the Section of Molecular Cytology...

...career landscape and there is insufficient (or no) emphasis on developing transferable skills to enable pursuit...recent discussions of this apparently worsening crisis recently appeared in eLife: "A Fair Deal for PhD...

...Saftic) to educational gifs created by the Amoeba Sisters. Here’s one example of how they “bridge the jargon...jargon gap using gifs”: https://amoebasisters.tumblr.com/post/139944465237/lactose-around-we-can-make-...

...lectures targeting three broad concepts: the genetic basis of disease, cancer, and viral infections. An interactive...science, but they didn’t seem to mind! Attendees consistently mentioned how pleased they were to learn about...

...you're transforming a plasmid with ampicillin resistance.  After a plasmid is introduced to bacteria, ...culture without antibiotics to give the antibiotic resistance gene time to be expressed.  However, you can ...

...perturbations that may play significant roles in cancer pathogenesis. Their work culminated in the first functional...lentiviral constructs using the High-Throughput Mutagenesis and Molecular Barcoding (HiTMMoB) technique (...

...genes as drivers of peritoneal mesothelioma tumorigenesis. They plan to use CRISPR and lentiviral plasmids...peritoneal mesothelioma cells lines and assess their tumorigenesis. These experiments will serve to validate these...

...they also went in search of genes involved in resistance to the cancer drug vemurafenib. Enriched libraries... screen for genes involved in bacterial toxin resistance, identifying 4 previously unknown genes.  “With...

...Optogenetic Tools Like pDawn  Making cassava plants resistant to cassava mosaic virus Dr. Fiona Robertson, University... cry genes from the soil bacterium Bacillus thuringiensis and analysing the insecticidal potential of ...

...microtubules. Tanenbaum et al. then tested their hypothesis that the lower expression levels of SunTag constructs...suitability in your experiments on a protein-by-protein basis. However, since many of Addgene’s SunTag plasmids...

...and undigested samples of the clones on an electrophoresis gel in order to differentiate which clone had...Once we had gotten our results back from the electrophoresis, my manager and I were unsure if there were...

...  Joining the Addgene crew!  This pattern was consistent: everyone I spoke with at Addgene, no matter ...presentation.  My first time in the lab! I got to assist production on a protocol video. Don't worry - no...

...neuronal circuit tracing.     Marcus Leiwe is an assistant professor at Kyushu University. He has a particular...resolution using chemical cocktails and computational analysis." Cell 157.3 (2014): 726-739. PubMed PMID: 24746791...

... dynamic process which involves autophagosome synthesis, delivery of materials to be degraded to the lysosome...mammalian dual-emission autophagy biosensor. Its resistance to lysosomal proteases is key to its pH-dependent...