Paul.xu: Created page with ". Antibody Library Construction Total RNAs were extracted from the spleens of hyperimmunized chickens to synthesize cDNA copies. The copies were used as templates to amplify t..."

2019-10-07T12:35:15Z

Created page with ". Antibody Library Construction Total RNAs were extracted from the spleens of hyperimmunized chickens to synthesize cDNA copies. The copies were used as templates to amplify t..."

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. Antibody Library Construction Total RNAs were extracted from the spleens of hyperimmunized chickens to synthesize cDNA copies. The copies were used as templates to amplify the VL and VH fragments of immunoglobulins using PCR. The amplified products were joined and extended by overlapping PCR to form single-chain variable fragments (scFv) approximately 750 bps long and containing a short or long peptide linker (data not shown). The recombinant PCR products were ligated into pComb3X vectors, cloned, and electroporated into Escherichia coli (E. coli) to construct two scFv antibody libraries. The resulting libraries were estimated to contain 9.0 × 108 phages with short linkers (scFv-S) and 8.4 × 108 phages with long linkers (scFv-L). After infecting E. coli with an M13 helper phage, the resulting phages were utilized for subsequent biopanning

Chicken Antibody Cloning - Revision history

Paul.xu: Created page with ". Antibody Library Construction Total RNAs were extracted from the spleens of hyperimmunized chickens to synthesize cDNA copies. The copies were used as templates to amplify t..."