Method systems and reagents for DNA sequencing

Details Method systems and reagents for DNA sequencing Method systems and reagents for DNA sequencing

1992-01-16

1994-04-26

Yarbrough, Amelia Burgess

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435291, 435808, 356344, 422 8207, 422 8208, 436171, 436172, 935 85, C12Q 168

Patent

active

053066189

ABSTRACT:
A DNA sequencing system and method are described to detect the presence of radiant energy emitted from different excited reporter dye-labeled species (DNA fragments) following separation in time and/or space, and the identity of the species which emit radiant energy closely spaced in wavelength. Functions of the emitted energy are obtained which vary over the wavelengths of the closely spaced spectra in different senses and the functions ratioed, whereby the ratio is indicative of the identity of the DNA fragments.
The emitting portion of the reporter-labeled DNA fragment is preferably one of a family of fluorescent dyes based on 9-carboxyethyl-6-hydroxy-3-oxo-3H-xanthene. These xanthene dyes are covalently attached to the DNA fragments through the carboxylic acid functionality, preferably via an amide linkage. The dyes may be protected by including an alkoxy group at the 9-position. A spacer may be inserted between the dye and the amine. The fluorescent dye preferably is attached to the DNA chain terminators and provides many advantages. Thus only DNA sequencing fragments resulting from bona fide termination events will carry a reporter. The DNA sequencing may also be labeled using the xanthene dyes which have general utility as fluorescent labels. Also acyclonucleoside triphosphates are described as being useful as chain terminators in DNA sequencing using a modification of the Sanger method.

REFERENCES:
patent: 3939350 (1976-02-01), Kronick et al.
patent: 4499052 (1985-02-01), Fulwyler
patent: 4748129 (1988-05-01), Chang et al.
Hendrickson et al., "Organic Chemistry", McGraw-Hill Book Company (1970) pp. 246-256.
Soini et al., "Fluoroimmunoassay: Present Status and Key Problems", Clin. Chem. 25/3, (1979) 353-361.
Z. G. Chidgeavadze and R. Sh. Beabealashvilli, "2', 3'-Dideoxy.varies.3' aminonucleoside 5'-triphosphates are the terminators of DNA synthesis catalyzed by DNA polymerases", 1984, pp. 1671-1686, Nucleic Acids Research vol. 12 No. 3.
A. Krayevsky, M. Kukhanova, A. Atrazhev, A. Azhayev, Z. Chidgeavadze, and R. Beabealashvilli, "2', 3'-Dideoxy-3'-aminonucleoside 5'-triphosphates are the terminators of DNA synthesis catalyzed by DNA polymerases", 1984, pp. 283-284, Nucleic Acids Research Symposium Series No. 14.
Robert S. Beabealashvilli, Andrei V. Scamrov, Tamara V. Kutateladze, Alexander M. Mazo, Alexander A. Krayevsky, and Marina K. Kukhanova, "Nucleoside 5'-triphosphates modified at sugar residues as substrates for calf thymus terminal deoxynucleotidyl transferase and for AMV reverse transcriptase", 1986, pp. 136-144.
Zurab G. Chidgeavadze, Robert Sh. Beabealashvilli, Alexander A. Krayevsky, and Marina K. Kukhanova, "Nucleoside 5'-triphosphates with modified sugars as substrates for DNA polymerases", 19896, pp. 145-152.
Naimski, Bierzynski, and Fikus; Anal. Biochem. 106, 471-475, 1980.
S. Folestad et al., Anal. Chem. 54, 925-979 1982.
A. M. Maxam, et al., Meth. in Enzym. 65 499-559 (1980).
F. Sanger et al., Proc. Nat. Acad,. Sci. USA 74 5463-5467 (1977).
L. M. Smith, Nucleic Acids Research, 13 2399-2412 (1985).

Affiliated with

Also associated with

Rating

Method systems and reagents for DNA sequencing does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Method systems and reagents for DNA sequencing, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Method systems and reagents for DNA sequencing will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-1711098