Abstract:
:A novel and simple method for detection of mutations in DNA oligonucleotides using a double-stranded DNA specific dye (SYBR Green I) is reported. The SYBR Green I is bound specifically with a duplex DNA oligonucleotide (it tercalation). This intercalation induces fluorescent emission at 525 nm with excitation at 494 nm. The fluorescence intensity of mismatched oligonucleotides (40-mer) decreases (by more than 13%) in comparison with the perfectly matched oligonucleotides. Moreover, fluorescence measurement of the SYBR Green I can distinguish various types of single-base mismatches, except for the T-G terminal mismatch. The addition of 20% (v/v) formamide, however, to an oligonucleotide solution improved the sensitivity of detection and also enabled the detection of the T-G terminal-mismatch. This detection method requires only a normal fluorescence spectrophotometer, an inexpensive dye and just 50 pmol of sample DNA.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Maruyama T,Takata T,Ichinose H,Park LC,Kamaiya N,Goto Mdoi
10.1023/a:1025661730518keywords:
subject
Has Abstractpub_date
2003-10-01 00:00:00pages
1637-41issue
19eissn
0141-5492issn
1573-6776journal_volume
25pub_type
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