CTAB-urea method purifies RNA from melanin for cDNA microarray analysis.
Melanin represents a major problem for the study of melanoma by microarrays since it is retained during RNA extraction and inhibits the enzymatic reactions used for probe preparation. Here we report a new method for cleaning RNA from melanin, based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB)-urea for RNA precipitation. This method is easy to perform and has a low cost. Purified RNA is recovered with high quality and good yield. CTAB-urea treated RNA from highly pigmented melanoma cells can be successfully reverse transcribed and labeled to obtain probes which can be subsequently used in cDNA microarray experiments, giving consistent and reproducible results.
Lagonigro
M Stefania
MS
Istituto FIRC di Oncologia Molecolare, via Adamello 16, Milan, Italy.
De Cecco
Loris
L
Carninci
Piero
P
Di Stasi
Delia
D
Ranzani
Tiziana
T
Rodolfo
Monica
M
Gariboldi
Manuela
M
D001709:Biotechnology; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry*; D018076:DNA, Complementary / Q000032:analysis*; D003902:Detergents; D008543:Melanins / Q000737:chemistry*; D008545:Melanoma / Q000737:chemistry / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000302:isolation & purification*; D015203:Reproducibility of Results; D014508:Urea / Q000737:chemistry*
D002593:Cetrimonium Compounds; D018076:DNA, Complementary; D003902:Detergents; D008543:Melanins; D012313:RNA; D014508:Urea; D000077286:Cetrimonium