Atlas of Human Anatomy vol.I-Musculoskeletal System by R. D. Sinelnikov and Ludmila Aksenova PDF

By R. D. Sinelnikov and Ludmila Aksenova

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3. Gateway® LR Clonase™ II Plus enzyme mix (Invitrogen; Life Technologies, Grand Island, NY). 4. Genomic DNA from species of interest. 5. Primers and solutions for PCR amplification. 6. Thermal-cycler. 7. Standard reagents and associated buffers for conventional cloning by restriction enzyme digestion and ligation: Xba, XhoI, ligase, ethidium bromide-agarose gel reagents and running buffers. 8. Commercial maxi-prep kit for preparation of purified plasmid DNA. 9. Primers for DNA sequencing designed to confirm correct cloning: T3 primer.

Details for otic cup injections can be found in an excellent methods chapter by Freeman et al. [23]. Here we focus on otocyst stages (see Note 7). 3. Slowly lower the micropipette until the tip pierces the epithelium of the otocyst. In some cases, the moment the otocyst is punctured, it can be visualized as a sudden alleviation of resistance applied to the needle tip by the epithelium, causing the embryo to recoil upward. 4. Inject the DNA mixture into the otocyst cavity until it begins to enlarge (Fig.

The size of the microcarrier should be optimized for the particular application, cell types, etc. For example, nanoparticles are used in the improved method of neuron transfection in the cultured brain slices [61–63]. However, whether nanoparticles under similar conditions penetrate the cuticular plate of hair cells needs further investigation. Tungsten particles are less expensive, but can oxidize and may be toxic to the cells. It is not recommended to use tungsten particles instead of gold [64].

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