Abstract: Detection of variable nucleotide(s) is based on primer extension and incorporation of detectable nucleoside triphosphates. By selecting the detection step primers from the region immediately adjacent to the variable nucleotide, this variation can be detected after incorporation of as few as one nucleoside triphosphate. Labelled nucleoside triphosphates matching the variable nucleotide are added and the incorporation of a label into the detection step primer is measured. The selection of the detection step primer is important to the method according to this invention and is dependent on the nucleotide sequence of interest. The detection step primers are preferably selected so as to span the region immediately toward the 3' end from the variable nucleotide to be detected. The detection step primers can also be complementary to a sequence beginning several nucleotides removed from the variable nucleotide.
Type:
Grant
Filed:
December 2, 1993
Date of Patent:
January 11, 2000
Assignee:
Molecular Tool, Inc.
Inventors:
Hans E. Soderlund, Anne-Christine Syvanen
Abstract: The invention concerns a reagent composition that employs at least two different terminators of a nucleic acid template-dependent primer extension reaction to determine the identity of a nucleotide base at a specific position in a nucleic acid of interest. The invention also concerns an immobilized method for determining such identification. The invention may be used to determine the presence or absence of a specific nucleotide sequence in a sample. It may also be employed in determination of genotype and in the identification of different alleles.
Type:
Grant
Filed:
October 11, 1991
Date of Patent:
December 21, 1999
Assignee:
Molecular Tool, Inc.
Inventors:
Philip Goelet, Michael R. Knapp, Stephen Anderson
Abstract: The invention concerns a reagent composition that employs at least two different terminators of a nucleic acid template-dependent primer extension reaction to determine the identity of a nucleotide base at a specific position in a nucleic acid of interest. The invention also concerns the method for determining such identification. The invention may be used to determine the presence or absence of a specific nucleotide sequence in a sample. It may also be employed in determination of genotype and in the identification of different alleles.
Type:
Grant
Filed:
March 5, 1991
Date of Patent:
March 30, 1999
Assignee:
Molecular Tool, Inc.
Inventors:
Philip Goelet, Michael R. Knapp, Stephen Anderson
Abstract: Methods for the covalent, specific and reversible immobilization of nucleic acid molecules onto solid-phases by means of a reversible disulfide bond for nucleic acid molecule array preparation are described. These methods can be used to prepare reusable nucleic acid molecule arrays with high specificity and high efficiency.
Abstract: A device determines the genotype at selected loci within genetic material obtained from a biological sample. One or more data sets are formed and a set of probability distributions including at least one distribution is established. These distributions associate hypothetical reaction values with corresponding probabilities for each genotype of interest at the same locus or at different loci. The genotype is then determined based on these measures.
Abstract: A method is provided for determining the identity of a nucleotide at a preselected site in a nucleic acid molecule. The method involves the incorporation of a nucleoside triphosphate that is complementary to the nucleotide present at the preselected site onto the terminus of a primer molecule, and their subsequent ligation to a second oligonucleotide. The reaction is monitored by detecting a specific label attached to the reaction's solid phase or by detection in solution.
Type:
Grant
Filed:
August 9, 1996
Date of Patent:
October 21, 1997
Assignee:
Molecular Tool, Inc.
Inventors:
Theo Nikiforov, Jonathan Karn, Philip Goelet
Abstract: Synthetic nucleic acid molecules are non-covalently immobilized in the presence of a salt or cationic detergent on a hydrophilic polystyrene solid support containing an --OH, --C.dbd.O or --COOH hydrophilic group or on a glass solid support. The support is contacted with a solution having a pH of about 6 to about 8 containing the synthetic nucleic acid and the cationic detergent or salt. Preferably, the cationic detergent is 1-ethyl-3-(3'-dimethylaminopropyl)-1,3-carbodiimide hyrochloride at a concentration of about 30 mM to about 100 mM or octyldimethylamine hydrochloride at a concentration of about 50 mM to about 150 mM. The salt is preferably NaCl at a concentration of about 50 mM to about 250 mM. When the detergent is 1-ethyl-3-(3'-dimethylaminopropyl)-1,3-carbodiimide hyrochloride, the glass support or the hydrophilic polystyrene support is used. When NaCl or octyldimethylamine hydrochloride is used, the support is the hydrophilic polystyrene.
Abstract: A method for generating single-stranded nucleic acid molecules. The molecules contain nuclease resistant modified nucleotides, such that they are resistant to 5'.fwdarw.3' exonucleases.