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5x106 HEK 293T cells plated in to 150 mm dishes were transfected with 10µg of TS shRNA plasmid, pLKO.1 vector or pSMPUW-GFP, 5µg psPAX (lentiviral packaging plasmid) and 5µg pMD2.G (VSV-G envelope expressing plasmid) with Fugene transfection Reagent (Qiagen), according to manufacturer’s instructions. 48 h post-transfection, media containing the lentiviral particles were harvested and centrifuged at 3000 rpm at 4°C for 10 min. The supernatant was collected, mixed with 1/3rd volume of fresh media and 1:1000µl polybrene was added.
Copyright and License information: ©2021 Haritha, Nawab, Vijayakurup, Anto, Liju, Alex, Amrutha, Aiswarya, Swetha, Vinod, Sundaram, Guijarro, Herlevich, Krishna, Nestory, Bava, Sadasivan, Zajac-Kaye and Anto
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