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METHOD FOR LARGE SCALE cDNA CLONING AND SEQUENCING BY CIRCULATING SUBTRACTION
METHOD FOR LARGE SCALE cDNA CLONING AND SEQUENCING BY CIRCULATING SUBTRACTION
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机译:通过循环减法进行大规模cDNA克隆和测序的方法
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摘要
The present invention provides a method of cDNA sequencing by circulating subtraction, which comprises the steps of: (1) constructing the cDNA original libraries with high quality using the organism tissues as materials; (2) homogenizing the cDNA original libraries of step (1) according to the graded C0t value, thereby obtaining the homogenized cDNA libraries corresponding to the graded C0t value; (3) selecting and sequencing the clones from each of the homogenized cDNA libraries of the preceding step; (4) hybridizing and subtracting the homogenized cDNA libraries of the preceding step by using the sequenced cDNA clones; (5) repeating steps (3) and (4) 1-5,000 times. Further, a step of preliminary subtractive hybridization may be included in the method. By using said method, one can sequence cDNA of certain tissues or species efficiently and comprehensively.
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