Molecular Tool, Inc.

Orchid Biocomputer, Inc.

Theo T Nikiforov

Jonathan Karn

Philip Goelet

Michael R Knapp

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.

Ligase/polymerase-mediated genetic bit analysis of single nucleotide

Ligase/polymerase mediated genetic bit analysis of single nucleotide

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. After immobilization, the support containing the immobilized nucleic acid may be washed with an aqueous solution containing a non-ionic detergent. The immobilized nucleic acid may be used in nucleic acid hybridization assays, nucleic acid sequencing and in analysis of genomic polymorphisms.

Method for immobilizing nucleic acid molecules

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.

Growing community of inventors

Baltimore, MD, United States of America

Theo T Nikiforov