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Strategy used for construction of naïve scFv-phage display libraries. Total RNA is isolated from B-lymphocytes from non-immunized healthy donors. Then cDNA is synthesized from the isolated RNA using reverse transcriptase enzyme. Then the repertoire of the VH and VL genes is amplified from the cDNA using forward and reverse primers hybridizing to the variable domains. scFvs are constructed and cloned into phagemid vector and a naïve phage library of 108-1010 is usually generated.
