RNA isolation from Xenopus inner ear sensory endorgans for transcriptional profiling and molecular cloning.
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 for experimental manipulation during animal development. However, many procedures for analyzing gene expression in the peripheral auditory and vestibular systems mandate the ability to isolate intact RNA from inner ear tissue. Methods presented here facilitate preparation of high quality inner ear RNA from larval and post-metamorphic Xenopus specimens that can be used for a variety of purposes. We demonstrate that RNA isolated with these protocols is suitable for microarray analysis of inner ear organs, and for cloning of large transcripts, such as those for ion channels. Genetic sequences cloned with these procedures can be used for transient transfection of Xenopus kidney cell lines with GFP fusion constructs.
Trujillo-Provencio
Casilda
C
Biology Department, New Mexico State University, Las Cruces, NM, USA.
Powers
TuShun R
TR
Sultemeier
David R
DR
Serrano
Elba E
EE
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D046169:Dermoscopy; D007758:Ear, Inner / Q000294:innervation / Q000378:metabolism*; D020869:Gene Expression Profiling; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011984:Sensory Receptor Cells / Q000378:metabolism*; D014982:Xenopus laevis
D012313:RNA