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Cloning strategy.

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posted on 05.02.2021, 21:06 by Virginia R. Pearson, Jens B. Bosse, Orkide O. Koyuncu, Julian Scherer, Cristhian Toruno, Rosann Robinson, Lisa M. Abegglen, Joshua D. Schiffman, Lynn W. Enquist, Glenn F. Rall

The T antigen-encoding region was amplified from elephant BW1 MaleNOD1, then inserted into the plasmid pEGFP-N1 (accession #U55762). DH5A competent bacteria were transformed and grown on kanamycin-resistant LB and amplified in a PCR reaction. PCR products were analyzed for expected band size of 2265 bp. Appropriate size gel bands were sequenced with standard primers (CMV forward and SV40 reverse to verify the presence and integrity of the AeIPyV-1 early region sequence.