...coli B and W strains. The optimized plasmid curing method was much simpler and took only ~32 hours instead...serve to interrupt genes (or gene clusters) in metabolic engineering. So far, we’ve used the MUCICAT system...when compared to the industrial strain. In our metabolic engineering case study of N-acetylglucosamine ...Multiple induction systems to choose from. Ready for metabolic engineering with improved cargo dedicated to gene...from our CRISPR topic overview Find other CRISPR methods for bacteria Find all articles about microbiology...

...What is Rett Syndrome? Caused by a mutation in the methyl-CpG binding protein 2 (MECP2) gene, Rett Syndrome...et al., 1999). We now know that MECP2 binds to methylated CpG dinucleotides and CAC trinucleotides during...caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185–188 . https.../13810 Amir RE, Zoghbi HY (2000) Rett syndrome: Methyl-CpG-binding protein 2 mutations and phenotype-genotype...after Intracisternal Delivery. Molecular Therapy - Methods & Clinical Development 5:106–115 . https://doi....

...It is also worth noting that plasmids can be symmetrically incompatible, meaning that both plasmids are... from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher... higher rate than the coresident plasmid. Asymmetric plasmid loss occurs for several reasons, including...high copy incompatible plasmids that lead to asymmetrical plasmid loss of the usually large, low copy ...Suzuki K, Yoshida K (2002) A novel plasmid curing method using incompatibility of plant pathogenic Ti plasmids...

...integrins. Six of the 11 IPI antibodies contain RGD-mimetic sequences that imitate this motif and compete with... unconjugated and recommended for use in flow cytometry. Based on the established applications of the ...resulting in cells that elongated and morphed asymmetrically (Jo et al., 2022). By contrast, LVDP-binding...conferred less force, leading cells to spread into symmetrical circles. While elucidating the role of tension...Addgene's Antibody Guide Antibodies 101: Flow Cytometry Antibodies 101: Introduction to Antibodies...

...non-conservative and conservative. The conservative methods are characterized by use of a homologous donor ...break-induced repair (BIR). The non-conservative method is single strand annealing (SSA), an error prone...Alterations. Edited by Doudna J and Sontheimer EJ. Methods in Enzymology Vol. 546; (2015). PubMed. Optimization...catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Richardson CD, et al. Nat Biotech....Genome Engineering. 2017.  Easi-CRISPR: a robust method for one-step generation of mice carrying conditional...

...minutes, they are perfect for molecular cloning and metabolic engineering.  The E. coli genome has an extensive...address many areas of microbial research like metabolism. Transport, biofilms, and much more. Because ..., breakthroughs in gene modification tools and methods such as CRISPR have vastly improved scientists’...variation between different plant species and metabolic properties affecting adaptations.  Arabidopsis...doi.org/10.1159/000490898 Gibbs, R., Weinstock, G., Metzker, M. et al. Genome sequence of the Brown Norway ...

...is out of your hands, it’s helpful to focus on something controllable, like experiments (and their controls...use a preadsorption control to confirm. In this method, the primary antibody is incubated with its antigen...that the target protein was measured on.    Flow cytometry In addition to the controls listed above, viability...compensation controls are recommended for flow cytometry. A viability control allows you to detect dead...fluorophore and are critical to analyze multiplex flow cytometry experiments. We hope that this blog post will ...

... using CRISPR is easier than more conventional methods. Time and money savings come from the fact that...mouse editing when compared to more conventional methods. First, one can work with almost any strains of... be $50,000. Finally, compared to conventional methods of genome engineering, CRISPR is much more efficient...2016) GONAD: A Novel CRISPR/Cas9 Genome Editing Method that Does Not Require Ex Vivo Handling of Embryos...101: Mammalian Expression Systems and Delivery Methods Learn about Mouse Work on Exercise and Depression...

...create stable transgenic cell lines. The traditional method of making stable cell lines is to transfect, select...piggyBac™ gene editing system compare with other methods of gene editing? There are primarily two other .../loxP) to remove the donor cassette. While this method can be effective, a loxP site is left behind in...Pluripotent Stem Cells with Seamless Gene Editing.,” Methods Mol. Biol., Apr. 2016. PubMed PMID: 27075976. 4.... J. Adams, “A high-throughput splinkerette-PCR method for the isolation and sequencing of retroviral ...

...protein labeling with trimethoprim conjugates: A flexible chemical tag. Nature Methods, 2(4), Article 4. ... Cohen lab used HaloTag for their “ticker tape” method for intracellular recordings, switching between...H., Vogel, H., & Johnsson, K. (2003). A general method for the covalent labeling of fusion proteins with.... https://doi.org/10.1038/nmeth749 Strebinger, D., Frangieh, C. J., Friedrich, M. J., Faure, G., Macrae...

...concentration. Three methods are broadly used for virus titration: the p24 ELISA dosing method detects the expression...expression of viral capsid antigen p24, the FACS dosing method correlates viral titre to the number of cells transduced... uses qPCR to quantify viral RNA. All of these methods have been described in a previous Addgene blog ...with the virus in order to aid viral binding. This method, called spinoculation, has been found very effective...

... you can help ensure that you understand the methodology and can have confidence in the reagents and outcomes...is preferable to finding out you've been doing something all wrong or that a reagent has not been convincingly...feedback from experts. If someone crosses out something, find out why. Ask successful writers about their...exercise. I highly recommend running or biking or something that demands your attention. Eat well- a diet... help maintain your health. Have a hobby- do something that gets you away from thinking about the lab...

...commentary about the new work published in Cell Metabolism. But, they note, there is plenty more to explore...be important to identify the key proteins and metabolites that are dynamically secreted by muscle during... Skeletal muscle PGC-1α1 modulates kynurenine metabolism and mediates resilience to stress-induced depression...Muscle over mind. Moon HY, van Praag H. doi: 0.1016/j.cmet.2014.09.012. PMID: 25295784 Related Materials:...

...breakthrough in cloning technology with the Golden Gate method. For the first time, it was possible to quickly...tedious and time-consuming traditional cloning methods, Lohmann said. The key component of Golden Gate...their lab learned the hard way after the cloning method, which had been humming along at 99 percent efficiency... et al. "A One Pot, One Step, Precision Cloning Method with High Throughput Capability." PLoS ONE. 3(11...

...A general method to fine-tune fluorophores for live-cell and in vivo imaging. Nature Methods, 14(10), ...technically cell-permeable, its use in live cells can sometimes result in cellular toxicity due to the permeation...to small-molecule fluorescent labels. In Nature Methods (Vol. 19, Issue 2, pp. 149–158). Nature Research..., 987–994. https://doi.org/10.1038/nmeth.4403 Hayashi-Takanaka, Y., Stasevich, T. J., Kurumizaka, H., ...

...In complex metazoans, rapid cell division and large scale cell mobility are essential processes during...specificity or from the thymus to the lymph nodes. These methods gave researchers much more information regarding...allows imaging of living tissue up to about one millimeter in depth. Two low-energy photons (typically from... between several different immune cells. These methods allowed immunologists to develop a spatiotemporal...absorption properties of fluorescent proteins." Nature methods 8.5 (2011): 393-399. PubMed PMID: 21527931. PubMed...

...world of academia often emphasizes structured methodologies and rigorous research plans. However, amidst...in nature or visiting museums can be excellent methods to detach from the constant stream of thoughts ...PhD studies, where every aspect of research is meticulously planned, it's easy to get stuck in your everyday...of postdocs focusing on molecular biology and metabolism. He now guides fellow PhDs in their careers with...

...sense dopamine. Methods for measuring Dopamine In vivo microdialysis The gold standard method of measuring... liquid for quantitation. The advantage of this method is that it can be performed in a live awake animal...the probe into a specific brain region. While the method is powerful and offers immediate measurements in...

...hormones strigolactones By Fabienne Furt Current methods used to detect strigolactones (SLs) are costly,...that of the internal control (Fig 1). Because this method relies on the fluorescence ratio between the 2 ...approach was also used to build and validate a ratiometric fluorescent biosensor based on the parasitic ...CRISPR library from Alec Kimmelman targets mouse metabolic genes with ~6 gRNAs per gene.  CRISPR-Act3.0, ...

...analysis in Drosophila. Nature Methods 9:47–55 . https://doi.org/10.1038/nmeth.1800 Isaacman-Beck J, Paik ...Maimon G, Wilson RI, Clandinin TR (2019) SPARC: a method to genetically manipulate precise proportions of...-wide reconstruction of single neurons. Nature Methods 15:1033–1036 . https://doi.org/10.1038/s41592-018...