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Subtraction of cochlea cDNA (tester) was performed with the PCR-Select cDNA Subtraction Kit (Clontech). Driver cDNA consisted of a mixture of cDNAs from brain, kidney, liver, heart, skeletal muscle, placenta, stomach (mucosal layer) and psoas muscle. Tester and driver cDNAs were digested by RsaI; adaptors were ligated to the tester. Two rounds of subtractive hybridization were performed, followed by two suppression PCR reactions. Subtracted PCR products were ligated into a T/A vector (Invitrogen).
Nucleotide
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