Process for amplifying, detecting, and/or-cloning nucleic acid sequences
This patent describes a process for detecting the presence or absence of at least one specific nucleic acid sequence in a sample containing a nucleic acid or mixture of nucleic acids, or distinguishing between two different sequences in the sample. The sample is suspected of containing the sequence or sequences. The process comprises: (a) treating the sample with one oligonucleotide primer for each strand of each different specific sequence, under hybridizing conditions such that for each strand of each different sequence to which an oligonucleotide primer is hybridized an extension product of each primer is synthesized which is complementary to each nucleic acid strand. The primer or primers are selected so as to be sufficiently complementary to each strand of each specific sequence to hybridize so that the extension product is synthesized from one primer. When it is separated from its complement, it serves as a template for synthesis of the extension product of the other primer; (b) treating the sample under denaturing conditions to separate the primer extension products from their templates if the sequence or sequences to be detected are present; (c) treating the sample with oligonucleotide primers such that a primer extension product is synthesized using each of the single strands produced in step (b) as a template, resulting in amplification of the specific nucleic acid sequence or sequences if present; (d) adding to the product of step (c) a labeled oligonucleotide probe for each sequence detected capable of hybridizing to the sequence or a mutation; and (e) determining whether the hybridization has occurred.
- Assignee:
- Cetus Corp., Emeryville, CA
- Patent Number(s):
- US 4683195
- OSTI ID:
- 6081384
- Resource Relation:
- Patent File Date: Filed date 7 Feb 1986
- Country of Publication:
- United States
- Language:
- English
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Process for amplifying nucleic acid sequences
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Related Subjects
DNA-CLONING
DESIGN
GENE AMPLIFICATION
PROBES
OLIGONUCLEOTIDES
FRACTIONATION
BIOSYNTHESIS
DETECTION
DNA SEQUENCING
HYBRIDIZATION
MUTATIONS
NUCLEIC ACID DENATURATION
SAMPLING
CLONING
NUCLEIC ACIDS
ORGANIC COMPOUNDS
SEPARATION PROCESSES
STRUCTURAL CHEMICAL ANALYSIS
SYNTHESIS
550200* - Biochemistry
550400 - Genetics