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The pMD19-T-PHD3 plasmids were digested by Hind III and Xho I restriction enzymes, and the target fragments (full length PHD3 cDNAs) were isolated and purified. The pcDNA 3.1(+) eukaryotic expression vectors were also digested by Hind III and Xho I and then ligated into PHD3 cDNA with DNA Ligation Kit v.2.0. |
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Protocol tips |
The pMD19-T-PHD3 plasmids were digested by Hind III and Xho I restriction enzymes, and the target fragments (full length PHD3 cDNAs) were isolated and purified. The pcDNA 3.1(+) eukaryotic expression vectors were also digested by Hind III and Xho I and then ligated into PHD3 cDNA with DNA Ligation Kit v.2.0. |
Publication protocol
The pMD19-T-PHD3 plasmids were digested by Hind III and Xho I restriction enzymes, and the target fragments (full length PHD3 cDNAs) were isolated and purified. The pcDNA 3.1(+) eukaryotic expression vectors were also digested by Hind III and Xho I and then ligated into PHD3 cDNA with DNA Ligation Kit v.2.0. The recombinant pcDNA 3.1(+)-PHD3 was amplified in E. coli DH5α competent cells, and isolated with TaKaRa MiniBEST Plasmid Purification Kit v.2.0. The correct pcDNA 3.1(+)-PHD3 plasmid sequence was verified by restriction enzyme mapping and DNA sequencing.
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