Real-time PCR products were directly ligatedinto plasmid pGEM-T Easy Vector (Promega),and transformed into E. coli JM109 High Effi-ciency competent cells (Promega) following themanufacturer’s protocols. White bacterial coloniescontaining plasmids with inserts were selected andplasmid DNA was purified using the WizardPlusSV miniprep DNA purification system (Promega).Purified plasmid concentrations were adjusted to20 ng µL−1 for sequencing. Sequencing was carriedout by the DNAseq sequencing service, Universityof Dundee, Scotland.
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