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Pooled PCR units were examined by agarose gel electrophoresis, Purified using the QIAquick gel extraction kit (Qiagen, santa Clarita, ca,bridestowe bear price, the united states) And ligated in the vector pCR2.1 (TOPOTA cloning, Invitrogen) or a pSCA (StrataClone PCR cloning, Stratagene, la Jollos angeles, florida, united states of america), after the manufacturers' recommended protocols. After improve of plasmids into host cells and blue screening, For each library inserts in 80 randomly picked colonies were proved using PCR with vectorspecific primers that flanked the cloning regions. The amplicons were digested with HaeIII and HhaI and sorted by restrictionfragment length polymorphism (RFLP) models on 3 MetaPhor agarose gels (BioWhittaker; Molecular purposes, Rockland,, to me, american).
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