NOVEL DNA SYNTHESIS TECHNOLOGY WITH 3’-BEADED OLIGO DNA AND DNA POLYMERASE

摘要

A method of synthesizing a desired DNA having a predetermined sequence, characterized by the steps of (a) preparing, based on the desired DNA, a plurality of 3'-biotin-immobilized oligo DNA having a fixed length; (b) preparing a starting DNA (1) which has a complementary sequence to a 3' end of a first immobilized oligo DNA (2) of the plurality of oligo DNA, and wherein the starting DNA (1) has a length shorter than the length of the first immobilized oligo DNA (2); (c) annealing the starting DNA (1) with the first immobilized oligo DNA (2), extending the starting DNA (1) to complement the first immobilized oligo DNA (2), thereby making a newly extended DNA (3); (d) denaturing a first double strand DNA (4) consisting of the newly extended DNA (3) and the first immobilized oligo DNA (2); (e) collecting the extended DNA (3) by removing the first immobilized oligo DNA (2) from the extended DNA (3); (f) annealing the collected, extended DNA (3) with a second immobilized oligo DNA (6) from the plurality of oligo DNA, further extending the extended DNA (3) to complement the second oligo DNA (6), thereby making a further extended DNA (7); (g) denaturing a second double stranded DNA (8) consisting of the second immobilized oligo DNA (5) and the further extended DNA (7); (h) collecting the further extended DNA (7) by removing the second immobilized oligo DNA (6) from the extended DNA (7); and (i) repeating steps (f) through (h) until the further extended DNA becomes the predetermined sequence thereby completing synthesis of the desired DNA.