Apparatus for identifying, classifying, or quantifying DNA seque

Details Apparatus for identifying, classifying, or quantifying DNA seque Apparatus for identifying, classifying, or quantifying DNA seque

1996-06-14

1999-10-26

Sisson, Bradley L.

Chemistry: molecular biology and microbiology

Apparatus

Including measuring or testing

436 94, 364496, 711100, C12M 100, G01N 128, H01L 2120

Patent

active

059726938

ABSTRACT:
This invention provides methods by which biologically derived DNA sequences in a mixed sample or in an arrayed single sequence clone can be determined and classified without sequencing. The methods make use of information on the presence of carefully chosen target subsequences, typically of length from 4 to 8 base pairs, and preferably the length between target subsequences in a sample DNA sequence together with DNA sequence databases containing lists of sequences likely to be present in the sample to determine a sample sequence. The preferred method uses restriction endonucleases to recognize target subsequences and cut the sample sequence. Then carefully chosen recognition moieties are ligated to the cut fragments, the fragments amplified, and the experimental observation made. Polymerase chain reaction (PCR) is the preferred method of amplification. Several alternative embodiments are described which capable of increased discrimination and which use TypeIIS restriction endonucleases, various capture moieties, or samples of specially synthesized cDNA. Another embodiment of the invention uses information on the presence or absence of carefully chosen target subsequences in a single sequence clone together with DNA sequence databases to determine the clone sequence. Computer implemented methods are provided to analyze the experimental results and to determine the sample sequences in question and to carefully choose target subsequences in order that experiments yield a maximum amount of information.

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