Specific triplex binding capacity of mixed base sequence duplex nucleic acids used for single-nucleotide polymorphism detection.
Specific base recognition and binding between native double-stranded DNA (dsDNA) and complementary single-stranded DNA (ssDNA) of mixed base sequence is presented. Third-strand binding, facilitated and stabilized by a DNA intercalator, YOYO-1, occurs within 5 min at room temperature. This triplex binding capability has been used to develop a homogeneous assay that accurately detects 1-, 2-, or 3-bp mutations or deletions in the dsDNA target. Every type of 1-bp mismatch can be identified. The assay can reliably distinguish homozygous from heterozygous polymerase chain reaction (PCR)-amplified genomic dsDNA, thus providing a highly sensitive clinical diagnostic assay.
Daksis
Jasmine I
JI
Ingeneus Research, Mississauga, Ontario, Canada L4Z 1V4. jasmine@ingeneus.net
Erikson
Glen H
GH
D020029:Base Pairing; D001583:Benzoxazoles; D002851:Chromatography, High Pressure Liquid; D004247:DNA; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D011806:Quinolinium Compounds; D013050:Spectrometry, Fluorescence; D013997:Time Factors
D001583:Benzoxazoles; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D011806:Quinolinium Compounds; C051802:triplex DNA; C075296:1,1'-((4,4,7,7-tetramethyl)-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-oxazole)-2-methylidene)quinolinium; D004247:DNA