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1st strand cDNA was primed with an oligo(dT) primer [ATGTGGCCTTTTTTTTTTTTTTTTT]; double-stranded cDNA was synthesized using specific 5' and 3' primers and amplified by PCR. The PCR product was digested with SfiI and size selection was performed to exclude fragments <1.5kb.The SfiI-digested PCR product was cloned into distinct DraIII sites of pME18S-FL3. XhoI sites just outside the DraIII sites can be used to isolate the cDNA insert. Libraries were constructed by oligo-capping method.
Nucleotide
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