A versatile method for the removal of melanin from ribonucleic acids in melanocytic cells.
Melanin pigments often co-purify during preparation of nucleic acids from cells or tissues of melanocytic origin. Contaminating melanin can severely impede subsequent analyses of RNA. We attempted to eliminate melanin in RNA preparations using selected gel matrices. We show here that co-purified melanin pigments can be largely eliminated from RNA samples after passing through polyacrylamide-based beads (Bio-Gel P-60). After isolation from the pigment-containing cells or tissues, RNA was subsequently processed through batch or column purification under acidic pH conditions. The resulting RNA was devoid of contaminating melanin pigments and amenable to molecular reactions such as polymerase chain reaction and cDNA synthesis by reverse transcriptase. Although the process results in some loss of input RNA, this purification procedure is simple, robust and can easily be adopted in any laboratory for the molecular analysis of RNA that requires removal of melanin contamination.
Satyamoorthy
K
K
The Wistar Institute, Philadelphia, PA 19104, USA. Herlynm@wistar.upenn.edu
Li
G
G
Van Belle
P A
PA
Elder
D E
DE
Herlyn
M
M
D000180:Acrylic Resins / Q000494:pharmacology; D015152:Blotting, Northern; D003582:Cytogenetics / Q000379:methods; D018076:DNA, Complementary / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008543:Melanins / Q000302:isolation & purification* / Q000378:metabolism; D008544:Melanocytes / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D000099166:Polyacrylamides
D000180:Acrylic Resins; D018076:DNA, Complementary; D008543:Melanins; D012313:RNA; D012194:RNA-Directed DNA Polymerase; D000099166:Polyacrylamides