By Bernd Sokolowski
This moment variation expands upon the former quantity with new and up-to-date chapters. Auditory and Vestibular learn: tools and Protocols, moment Edition courses readers via protocols on phone tradition, tissue engineering, nanotechnology, high-throughput screening, and physiology. Chapters on body structure disguise strategies that come with optical coherence tomography, patch clamping, and photostimulation of caged neurotransmitters. Written within the hugely profitable Methods in Molecular Biology series layout, chapters comprise introductions to their respective issues, lists of the required fabrics and reagents, step by step, without difficulty reproducible laboratory protocols, and pointers on troubleshooting and heading off identified pitfalls.
Authoritative and cutting-edge, Auditory and Vestibular learn: tools and Protocols, moment variation aims to make sure winning ends up in the additional research of this important field.
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Extra resources for Auditory and Vestibular Research: Methods and Protocols
If any of the DNA mixture remains, it can be stored at 4 °C or −20 °C, and reused. Turn off nitrogen tank. 11. Embryos should be left in a humidified incubator until the desired embryonic stage. Figure 5 shows evidence of gene transfection in sections through the inner ear 4 days after plasmid electroporation into the E3 otocyst. 12. Visualize GFP by immunofluorescence and miR-9 by in situ hybridization. Our lab has successfully collected transgeneexpressing basilar papillas 14 days after electroporation, although survival rates decrease dramatically beyond 3–4 days post-electroporation .
This is critical for successful electroporation. We have had success using a needle or forceps to pierce the membranes and deliver a small amount of Ringer’s solution beneath the amnion to separate it from the embryo before tearing it open, as described previously . 6. 5 Preparation of Injection Needle 1. 25 % fast green. Use a molar ratio of 2:1 for pTol2-GFPsd-miR(X)-sa and CAG-T2TP, respectively. Hereafter, we refer to this mix as plasmid DNA. A simplified version of each plasmid is shown schematically in Fig.
32. 33. 34. 35. 36. gamma-actin (ACTG1) mutations that cause DFNA20/26 hearing impairment. Hum Mol Genet 18:3075–3089 Kawashima Y, Géléoc GS, Kurima K et al (2011) Mechanotransduction in mouse inner ear hair cells requires transmembrane channellike genes. J Clin Invest 121:4796–4809 Diaz-Horta O, Subasioglu-Uzak A, Grati M et al (2014) FAM65B is a membrane-associated protein of hair cell stereocilia required for hearing. Proc Natl Acad Sci U S A 111: 9864–9868 Felgner PL, Gadek TR, Holm M et al (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure.