Fluorescent nucleotide analog hairpin formation for detection of nucleic ...

(12) United States Patent ao) Patent No.: us 6,451,530 Bi

Hawkins (45) Date of Patent: Sep. 17,2002

1 1 1 1 1 1 1

400 450 500 550

WAVELENGTH (NM)

FJ& I.

U.S. Patent

Sep. 17,2002

Sheet 2 of 3

US 6,451,530 Bl

nMyc gene

1 agctccgagc ccccgagcrg ggccacggag atgccgcctg agaacgagct gtggggcagc 61 ccggccgagg aggacgcgtrr cggccrgggg ggacngggtg gcctcacccc caacccggcc 121 atcctccagg accgcacgcg gagcggcttc tccgcccgcg agaagccgga gcgcgccgtg 181 agcgagaagc tgcagcacgg ccgcgggccg ccaaccgccg gttccaccgc ccagtccccg 241 ggagtrcggcg ccgccagccc tgcgggtcgc gggcacggcg gggccgcggg agccggccgc 301 gccggggccg cccrgcccgc cgagctcgcc cacccggccg ccgagtgcgt ggaccccgcc 361 gtggtctccc cccttcccgr gaacaagcgc gagccagcgc ccgcgcccgc agccccggcc 421 agtgccccgg cggcgggccc tgcggtcgcc tcgggggcgg gtattgccgc cccagccggg 481 gccccggggg tcgcccctcc gcgcccaggc ggccgccaga ccagcggcgg cgaccacaag 541 gccctcagta cctccggaga ggacaccctg agcgattcag atgatgaaga tgatgaagag 601 gaagatgaag aggaagaaac cgacgtggtc actgtggaga agcggcgttc ctcccccaac 661 accaaggctg rrcaccacatt caccatcact gtgcgtccca agaacgcagc cctgggtccc 721 gggagggctc agtccagcga gctgatcctc aaacgatgcc ttcccatcca ccagcagcac 781 aactatgccg ccccctcccc ctacgcggag agtgaggatg cacccccaca gaagaagata 841 aagagcgagg cgtccccacg tccgctcaag agtgtcaccc ccccaaaggc taaaggjictta 901 aacccccgaa actctaacrc caUaggacaqt gagcgtcgca gaaaccacaa caccctggag 961 cgccagcgcc gcaacgaccc tcggtccagc tttctcacgc tcagggacca cgtgccggag 1021 ttggtaaaga atgagaaggc cgccaaggtg gtcattttga aaaaggccac tgagtatgtc 1081 cactccctcc aggccgagga gcaccagctt ttgctggaaa aggaaaaatt gcaggcaaga 1141 cagcagcagt cgctaaagaa aattgaacac gctcggactt gctagacgct tctcaaaact 1201 ggacagtcac tgccactttg cacattttga tttttttttt aaacaaacat tgtgttgaca 1261 ttaagaatgt tggtttactt tcaa

Fig. 2

ScrF I Mse

Fok I EcoR II

Fok I BstN I

| | sk38-> |

ggaacaaataggatggatgacaUaataatccacctatcccaqtaqqaqaaatttataaaaqatqqataatcctqqgattaa

ccttqtttatcctacctactgtttattaggtggatagggtcatcctctttaaatattttctacctattaggaccctaatt

$11$ £ 8 8 £ I I £

1531 1590 1535 1590

1590

1590 1597

Bsm I Sau96 I

BstX I Ava II Nla IV Acc I

II II I

ataaaatagtaagaatgtatagccctaccagcattctggacataagacaaggaccaaaggaaccctttagagactatgta

t:at--ht:t:atcattcttacatatcgqqatqqtcqtaaqacctgtattctqttcctqgtttccttqqqaaatctctqatacat

$ £ I £1 *~ sk39 £1 £ I £

1628 1651 1659 1678

1631 1651

Fig. 3