Mutation analysis by mass spectrometry using photolytically...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

Reexamination Certificate

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C435S091100

Reexamination Certificate

active

10079043

ABSTRACT:
The invention relates to a method of a mass-spectrometric analysis of known mutation sites in the genome, such as single nucleotide polymorphisms (SNPs), using the method of restricted primer extension. The invention consists of the use of primers with a photocleavable linker. The linker creates a gap in a DNA strand which is almost the same size as a natural DNA building block (nucleoside). The linker forms a bridge over a base pair without inhibiting hybridization or enzymatic extension. However, the linker allows the primers to be cleaved after extension in order to obtain short DNA fragments which can be more easily detected on the mass spectrometer.

REFERENCES:
patent: 5547835 (1996-08-01), Koster
patent: 5830655 (1998-11-01), Monforte et al.
patent: 6043031 (2000-03-01), Köster et al.
patent: 6221601 (2001-04-01), Koster et al.
patent: 0 840 804 (1998-05-01), None
patent: WO 97/27325 (1997-07-01), None
Phillip Ordoukhanian and John-Stephen Taylor, Design and Synthesis of a Versatile Photocleavable DNA Building Block. Application to Phototriggered Hybridization, American Chemical Society 1995, vol. 117, pp. 9570-9571, St. Louis, Missouri.
Sascha Sauer, et al., A novel procedure for efficient genotyping of single nucleotide polymorphisms, Nucleic Acids Research, 2000, vol. 28, No. 5, Oxford University Press, Berlin-Dahlem, Germany.

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