Prestaining method as a useful tool for the agarose gel electrophoretic detection of polymerase chain reaction products with a fluorescent dye SYBR gold nucleic acid gel stain.
Three staining methods using SYBR Gold Nucleic Acid Gel Stain (SYBR Gold) as a fluorescent dye were evaluated for the agarose gel electrophoretic detection of DNA. The methods involve prestain, in-gel stain, and poststain methods. DNA markers and polymerase chain reaction (PCR) products obtained by minisatellite variant repeat-PCR (MVR-PCR) amplification in a D1S8 locus were used as model DNA and practical samples, respectively. Among the three methods tested under the usual electrophoretic conditions, a prestain method using a 10000-fold diluted SYBR Gold solution showed most excellent features regarding cost and rapidity to use with good stainability and resolution over loaded DNA amounts of about 98 ng to 300 ng. The prestain method was found to be applicable to the analysis of DNA in MVR-PCR products from a human hair root.
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
Emi
E
Suenaga
Hiroshi
H
Nakamura
D004396:Coloring Agents / Q000737:chemistry*; D004247:DNA / Q000096:biosynthesis / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D006197:Hair / Q000737:chemistry; D006801:Humans; D009930:Organic Chemicals / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D004396:Coloring Agents; D009930:Organic Chemicals; C474470:SYBR Gold nucleic acid gel stain; D004247:DNA