REACTION CONCENTRATE FOR DNA SEQUENCING WITH THERMOSTABLE DNA POLYMERASE.

摘要

The present invention provides a nucleotide sequencing reaction concentrate comprising: a) sufficient thermostable polymerase to provide from 100 to 500 I.U./mL; b) from 0.3 to 30 mM of a reducing agent selected from dithiothreitol and beta -mercaptoethanol; c) sufficient phosphate buffer to maintain a pH of about 7.5; d) at least 40% glycerol; e) from 15 to 70 mu M dGTP or daGTP; f) from 15 to 150 mu M dATP; g) from 15 to 150 mu M dTTP; h) from 10 to 18 mu M dCTP; and i) one member selected from the group consisting of: i) sufficient ddGTP to provide a ddGTP to dGTP or daGTP ratio of about 5; ii) sufficient ddATP to form a ddATP to dATP ratio of about 33; iii) ddTTP in a sufficient amount to form a ddTTP to dTTP ratio of about 28; and iv) ddCTP in a sufficient amount to form a ddCTP to dCTP ratio of about 22.