METHOD FOR LARGE SCALE cDNA CLONING AND SEQUENCING BY CIRCULATING SUBTRACTION

摘要

The present invention provides a method for cDNA sequencing by circulating subtraction, which comprises the steps of: (1) constructing cDNA original libraries with high quality using the tissue of organism as material; (2) according to the grade Cot value assumed, homogenising the cDNA original libraries of step (1) to obtain the cDNA homogeneous libraries; (3) from the cDNA homogeneous libraries of preceding step, choosing clones to sequence DNA; (4) using the cDNA clones which have been sequenced, hybrizing and subtracting the cDNA homogeneous libraries of preceeding step; (5) repeating step (3) and (4) for 1-5000 times. The method can be added a step of preliminary subtractive hybridisation. The method can sequence the cDNA of certain tissue or species effectively and comprehensively.