Construction of pcDNA5/RT/TO-NPAT plasmid (for conditional stable cell line production)

NPAT cDNA was amplified by PCR using a plasmid from Origene (RC220768) as a template. The PCR products were then extracted and purified before being ligated into the pGEM-T Easy vector according to manufacturer instructions. The full NPAT cDNA was excised from the pGEM-T Easy vector and cloned into the pcDNA5/RT/TO vector using the NotI restriction site. Plasmid sequencing was performed to confirm successful orientation and validity of full NPAT sequence.