Optimized methodology for sequential extraction of RNA and protein from small human skin biopsies.
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.
Berglund
Susanne R
SR
Department of Radiation Oncology, UC Davis Cancer Center, Sacramento, California 95817, USA.
Schwietert
Chad W
CW
Jones
Angela A
AA
Stern
Robin L
RL
Lehmann
Joerg
J
Goldberg
Zelanna
Z
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
D011506:Proteins; D012313:RNA