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. 2023 Apr 13;11(4):833. doi: 10.3390/vaccines11040833

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Construction of the pAK8-P1 phagemid. (A) Colony PCR with universal M13 primers: A 611 bp product is amplified in bacteria transformed with pAK8-P1 phagemid (lanes 1 to 3). Colony PCR on bacteria containing the pAK8 phagemid with unrelated inserts resulted in a 752 bp product (positive control for M13 phagemid, lane 4), and no amplification was observed in non-transformed bacteria (negative control, lane 5). (B) Restriction site mapping: Undigested phagemid was seen in lane 1. Digestion of pAK8 phagemid with EcoRI and HindIII resulted in 2560- and 3384-nucleotide DNA fragments, indicating the P1 gene’s insertion into the pAK8 phagemid, according to SnapGene analysis.