Method for the amplification of a base sequence

Details Method for the amplification of a base sequence Method for the amplification of a base sequence

1996-07-10

1998-06-23

Horlick, Kenneth R.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 6, 536 243, 536 2532, C12P 1934, C12Q 168, C07H 2104, C07H 2100

Patent

active

057704082

ABSTRACT:
The present invention relates to a method for the amplification of a base sequence A1A2 consisting of two successive base sequences, A1 and A2, in a single strand polynucleotide as an object to be amplified characterized in that said method comprising at least (1) hybridizing a probe B consisting of a polynucleotide comprising the base sequence B1 complementary to said base sequence A1, a polynucleotide comprising the base sequence B2 complementary to said base sequence A2 and a linkage portion linking the two polynucleotide via itself, with the base sequence A1A2 of said polynucleotide, as the object to be amplified, to form a hybrid, (2) ligating the 5'-end of the base sequence B1 in said probe B to the 3'-end of the base sequence B2 in the same with the aid of ligase, (3) heat denaturating the double strand formed said hybridization, (4) hybridizing said heat denaturated polynucleotide complex with another probe B or a probe A consisting of a polynucleotide comprising said base sequence A1, a polynucleotide comprising said base sequence A2 and a linkage portion linking the two polynucleotide via itself, and (5) ligating 5'-end of the base sequence B1 in said probe B to the 3'-end of the base sequence B2 in the same, or 3'-end of the base sequence A1 in said probe A to the 5'-end of the base sequence A2 in the same with the aid of ligase.

REFERENCES:
patent: 4914210 (1990-04-01), Levenson et al.
Nilsson et al. Padlock Probes: Circularizing Oligonucleotides for Localized DNA Detection. Science, vol. 265, pp. 2085-2088, 1994.

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