Shot-gun sequencing and amplification without cloning
Patent 6521428 Issued on February 18, 2003. Estimated Expiration Date: 
November 4, 2019. Estimated Expiration Date is calculated based on simple USPTO term provisions. It does not account for terminal disclaimers, term adjustments, failure to pay maintenance fees, or other factors which might affect the term of a patent.
November 4, 2019. Estimated Expiration Date is calculated based on simple USPTO term provisions. It does not account for terminal disclaimers, term adjustments, failure to pay maintenance fees, or other factors which might affect the term of a patent.Patent ReferencesDNA sequencing by stepwise ligation and cleavage Island hopping--a method to sequence rapidly very large fragments of DNA Method of DNA base sequence determination Patent #: 5935794 InventorApplicationNo. 434761 filed on 11/04/1999US Classes:435/91.2, Acellular exponential or geometric amplification (e.g., PCR, etc.)435/6, Involving nucleic acid435/91.1, Polynucleotide (e.g., nucleic acid, oligonucleotide, etc.)436/94, Saccharide (e.g., DNA, etc.)536/23.1, DNA or RNA fragments or modified forms thereof (e.g., genes, etc.)536/24.3, Probes for detection of specific nucleotide sequences or primers for the synthesis of DNA or RNA536/24.33PrimersExaminersPrimary: Whisenant, EthanAssistant: Lu, Frank Attorney, Agent or FirmInternational ClassesC12P 019/34C12Q 001/68 G01N 033/00 C07H 021/02 C07H 021/04 AbstractDisclosed is a method for sequencing and amplifying nucleic acid templates wherein a degenerate primer with a fixed sequence region and a random sequence region is utilized. By determining the statistical expectancy of the fixed sequence in the nucleic acid template, this determines the average length of a nucleic acid template that can be sequenced. During the annealing of such a primer with the nucleic acid template, the fixed sequence determines where the complete primer binds by binding to its complementary sequence on the nucleic acid template. The random sequence regions of the primers make it possible for the presence of a unique sequence adjacent to the fixed sequence to be present, thus providing a primer with full complementarity with the nucleic acid template. Thus, this procedure is able to provide a full-length primer with a fully complementary sequence capable of binding statistically once within an expected length of the nucleic acid template, even though the sequence of the template is unknown. The method can also be adopted for use in PCR amplification of a nucleic acid template.Other References
Field of SearchInvolving nucleic acidPolynucleotide (e.g., nucleic acid, oligonucleotide, etc.) Acellular exponential or geometric amplification (e.g., PCR, etc.) ENZYME (E.G., LIGASES (6. ), ETC.), PROENZYME; COMPOSITIONS THEREOF; PROCESS FOR PREPARING, ACTIVATING, INHIBITING, SEPARATING, OR PURIFYING ENZYMES Saccharide (e.g., DNA, etc.) DNA or RNA fragments or modified forms thereof (e.g., genes, etc.) Probes for detection of specific nucleotide sequences or primers for the synthesis of DNA or RNA Primers Synthesis of polynucleotides or oligonucleotides |
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