RNA Extraction from Xenopus Auditory and Vestibular Organs for Molecular Cloning and Expression Profiling with RNA-Seq and Microarrays

摘要: The amphibian Xenopus offers a unique model system for uncovering the genetic basis of auditory and vestibular function in an organism that is well-suited experimental manipulation during animal development. However, many procedures analyzing gene expression peripheral systems mandate ability to isolate intact RNA from inner ear tissue. Methods presented here facilitate preparation high-quality larval post-metamorphic specimens can be used variety purposes. We demonstrate isolated with these protocols suitable microarray analysis Illumina-Solexa sequencing (RNA-Seq) organs, cloning large transcripts, such as those ion channels. Genetic sequences cloned transient transfection kidney cell lines fluorescent protein fusion constructs.