title: Optimized methodology for sequential extraction of RNA and protein from small human skin biopsies.
abstract: Current translational human studies are moving in the direction of concurrent genomic and proteomic analysis using small clinical samples. Skin tissue, although easily accessible, is difficult to process owing to its natural resistance to mechanical shearing and high levels of RNases and proteases. Currently, these complications result in degraded RNA samples with variable yield. We have developed a method of sequential extraction of high quality RNA and protein from a single 3 mm full thickness skin punch biopsy. This method yields 1-2 microg of RNA and 150 microg of protein, which is usable in many sensitive downstream applications including microarray, quantitative real-time PCR, two-dimensional gel electrophoresis and Western blot analysis.
authors.affiliation: Department of Radiation Oncology, UC Davis Cancer Center, Sacramento, California 95817, USA.
authors.forename: Susanne R
authors.initials: SR
authors.lastname: Berglund
authors.forename: Chad W
authors.initials: CW
authors.lastname: Schwietert
authors.forename: Angela A
authors.initials: AA
authors.lastname: Jones
authors.forename: Robin L
authors.initials: RL
authors.lastname: Stern
authors.forename: Joerg
authors.initials: J
authors.lastname: Lehmann
authors.forename: Zelanna
authors.initials: Z
authors.lastname: Goldberg
mesh_terms: D001671:Biochemistry / Q000379:methods*; D001706:Biopsy; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D006801:Humans; D011506:Proteins / Q000302:isolation & purification*; D012313:RNA / Q000302:isolation & purification*; D012867:Skin / Q000737:chemistry* / Q000473:pathology
chemical_list: D011506:Proteins; D012313:RNA