Lipofectamine® 2000 Transfection Reagent

Protocol tips

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	Add 1:1 ratio of DNA to diluted Lipofectamine® 2000 Reagent and Incubate for 5 minutes at room temperature. Incubate cells for 1–3 days at 37°C. Then analyze transfected cells

Protocol tips
Add 1:1 ratio of DNA to diluted Lipofectamine® 2000 Reagent and Incubate for 5 minutes at room temperature. Incubate cells for 1–3 days at 37°C. Then analyze transfected cells

Publication protocol

The human AEG-1/MTDH siRNA oligonucleotides (Sequence 1: sense 5′-GCAGCAAGGCAGTCTTTAAG T-3′, antisense 5′-ACTTAAAGACTGCCTTGCTGC-3′; Sequence 2: sense 5′-GUUACCACCGAGCAACUUAdT dT-3′, antisense 5′-UAAGUUGCUCGGUGGUAACdT dT-3′), Beclin-1 (BECN1) siRNA oligonucleotides (Sequence 1: sense 5′-AAGAUUGAAGACACAGGAG GC-3′, antisense 5′-GCCUCCUGUGUCUUCAAUCU U-3′; Sequence 2: sense 5′-CAGUUUGGCACAAUCAA UA-3′, antisense 5′-UAUUGAUUGUGCCAAACUG-3′) and the rat AEG-1/MTDH siRNA oligonucleotides (Sequence 1: sense 5′-CAACAGCGUAAACGUGAUA TT-3′, antisense 5′-UAUCACGUUUACGCUGUUGTT-3′; Sequence 2: sense 5′-CACCGAGCAACUUACAACU TT-3′, antisense 5′-AGUUGUAAGUUGCUCGGUG TT-3′; Sequence 3: sense 5′-GGGAAGGAAUUGGAGU GAUTT-3′, antisense 5′-AUCACUCCAAUUCCUUCCC TT-3′) are synthesized by GenePharma, Inc. (Pudong, Shanghai). A universal negative control siRNA is used. We perform siRNA transfection using Lipofectamine™ 2000 (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. Briefly, the cells that reached 50% confluence are transfected with serum-free DMEM medium containing 50 nmol/l scrambled siRNA, AEG-1, or BECN1 siRNA for 6 hours, followed by recovery in medium containing serum and other special treatments as necessary.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Lipofectamine® 2000 Transfection Reagent below.

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