Compositions and methods for inhibiting terminal transferase activity
AssigneeIbis Biosciences, Inc.
InventorsSherilynn Manalili Wheeler, James C. Hannis, Steven A. Hofstadler
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Abstract
The present invention relates to systems and methods for amplifying nucleic acid. In particular, systems and methods are provided for inhibiting polymerase based terminal transferase activity within a polynucleotide amplification setting (e.g., polymerase chain reaction). In addition, systems and methods are provided for generating amplified products generated with polynucleotide amplification techniques having reduced 3′ non-templated nucleotide addition.
Claims28
A method for generating amplification products, comprising:
- a) combining boric acid and a reaction solution comprising a nucleic acid template, primers complementary to the nucleic acid template, deoxynucleotide triphosphates, a polymerase having terminal transferase activity and a buffer solution comprising KCl and Tris-HCl;
- and b) performing a polymerase chain (PCR) with said combined boric acid and said reaction solution such that amplification products are generated.
A method for generating amplification products, comprising:
- a) combining boric acid and a reaction solution comprising a nucleic acid template, primers complementary to the nucleic acid template, deoxynucleotide triphosphates, a polymerase having terminal transferase activity and a buffer solution comprising (NH)4SO4 and Tris-HCl;
- and b) performing a polymerase chain (PCR) with said combined boric acid and said reaction solution such that amplification products are generated.
A method for generating amplification products, comprising:
- a) combining boric acid and a reaction solution comprising a nucleic acid template, primers complementary to the nucleic acid template, deoxynucleotide triphosphates, a polymerase having terminal transferase activity wherein said boric acid is present at a final concentration of 100 mM to 200 mM in said reaction solution;
- and b) performing a polymerase chain (PCR) with said combined boric acid and said reaction solution such that amplification products are generated.
A method for generating amplification products, comprising:
- a) combining boric acid and a reaction solution comprising a nucleic acid template, primers complementary to the nucleic acid template, deoxynucleotide triphosphates, a polymerase having terminal transferase activity wherein said boric acid is present at a final concentration of 50 mM to 500 mM in said reaction solution;
- and b) performing a polymerase chain (PCR) with said combined boric acid and said reaction solution such that amplification products are generated.
Dependencies are inferred from the claim text.