EFFICIENT DNA-SEQUENCING ON MICROTITER PLATES USING DRIED REAGENTS AND AND BST DNA-POLYMERASE

Sequenase, Taq DNA polymerase and Bst DNA polymerase were tested for s equencing of DNA on microtiter plates using dried down reagents. Sever al parameters were investigated to expedite the drying process while m inimizing damage to the enzyme. Sequenase did not tolerate drying very well, and frequently generated sequences with weak signals and many s ites of premature termination. With Taq DNA polymerase it was possible to obtain sequences of good quality. However, there was considerable variation of results between experiments and between batches of microt iter plates. Bst DNA polymerase generated sequences of excellent quali ty. It was stable for more than a week in dried-down state at -20 degr ees C and at least overnight at room temperature. The method described here using Bst DNA polymerase is well suited for laboratory robots an d workstations that typically employ 96-well microtiter plates.