Enzyme composition for dna end repair, adenylation, phosphorylation
AssigneeThermo Fisher Scientific Baltics UAB
InventorsJudita LUBIENE, Arturas Berezniakovas, Arvydas Lubys
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Abstract
Enzyme compositions and their method of use that provide ready-to-use master mixtures. The compositions comprise a modified thermophilic DNA polymerase lacking 5′-3′ and 3′-5′ exonuclease activity premixed with T4 DNA polymerase, Klenow fragment and T4 polynucleotide kinase and all other necessary components, including reaction buffer and nucleoside triphosphates, required to perform DNA blunting, phosphorylation, and single nucleotide extension reactions in one tube and in two steps. Among other benefits, the mixture of different enzymes, buffers and nucleoside triphosphates is stable during prolonged storage.
Claims20
An enzyme composition comprising in a single container a buffer comprising nucleoside triphosphates and a plurality of enzymes separately capable of blunting, phosphorylating and adenylating DNA fragments, with the enzyme capable of adenylating DNA being a thermophilic modified DNA polymerase, and the composition being stable for at least one week at 4° C.
An enzyme composition comprising in a single container Tris-HCl, MgCl2, DTT, KCl, dATP, dCTP, dTTP, dGTP, ATP, Triton X-100, glycerol, NP 40, EDTA, Tween 20, T4 polynucleotide kinase, T4 DNA polymerase, Klenow fragment, and thermophilic modified DNA polymerase from Thermus brockianus or Thermus sp. fused to DNA-binding domain from Sso family, the composition contained in a single container and being storage stable.
Dependencies are inferred from the claim text.