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Last Updated: October 1, 2020

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Patent: 5,470,722

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Summary for Patent: 5,470,722
Title: Method for the amplification of unknown flanking DNA sequence
Abstract:A method that permits the rapid amplification of unknown DNA that flanks a known site, such that one can walk into an uncharacterized region of DNA. In this method, human genomic DNA is restriction enzyme digested and then ligated to a 5\' phosphorylated-oligonucleotide so that the 5\' end of each strand of genomic DNA is extended and phosphorylated. The phosphorylated-oligonucleotide is constructed to render 5\' end extensions that are complementary to the known sequence. Following denaturation and re-annealing under dilute conditions that promote intrastrand annealing and under high stringency, only those DNA strands containing the known sequence will form a stem-loop structure with a recessed and phosphorylated 5\' end, rendering a substrate for a subsequent heat-stable ligation reaction to another oligonucleotide. This second oligonucleotide is complementary to the sequence immediately adjacent to the phosphorylated-oligonucleotide high stringency annealing site. The heat-stable ligation reaction appends a known sequence to the DNA segments containing the two known contiguous DNA sequences used for oligonucleotide annealing. This heat-stable ligation of known sequence permits the subsequent highly specific amplification of the unknown flanking DNA.
Inventor(s): Jones; Douglas H. (Iowa City, IA)
Assignee: University of Iowa Research Foundation (Oakdale, IA)
Application Number:08/058,907
Patent Claims:see list of patent claims

Details for Patent 5,470,722

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Schering INTRON A interferon alfa-2b VIAL 103132 003 1986-06-04   Start Trial University of Iowa Research Foundation (Oakdale, IA) 2039-02-26 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 002 1986-06-04   Start Trial University of Iowa Research Foundation (Oakdale, IA) 2039-02-26 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 001 1986-06-04   Start Trial University of Iowa Research Foundation (Oakdale, IA) 2039-02-26 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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