Attractene Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 5.0 × 10^5 cells	Add 4.5 μL Attractene Transfection Reagent to the DNA solution. Incubate the cells under their normal growth conditions and change medium after 6h and continue to incubate until 48 h

Upstream tips
Seed 5.0 × 10^5 cells
Protocol tips
Add 4.5 μL Attractene Transfection Reagent to the DNA solution. Incubate the cells under their normal growth conditions and change medium after 6h and continue to incubate until 48 h

Publication protocol

HEK293T-CAPN5-p.R243L cells were transfected using Attractene Transfection Reagent (Qiagen). In each well of 6-well plates, 5.0 × 105 HEK293T-CAPN5-p.R243L cells were grown in 2 mL complete DMEM medium to 50 - 70% confluence. Growth medium was replaced with 2 mL of freshly prepared Opti-MEM medium containing 5% FBS, penicillin (100U/mL) and streptomycin (100 μg/mL) (complete Opti-MEM medium). Cells were then transfected with 3.0 μg shRNA plasmid using 4.5 μL Attractene Reagent. At 6 hours post-transfection, growth medium was replaced with fresh complete Opti-MEM medium. Cells were incubated for 48 hours.

The optimization protocols (varying pulse voltage, pulse width and pulse, as recommended by manufacturer) for transfecting adherent and suspension cells with Neon™electroporation system (Invitrogen; Carlsbad, CA) were performed with SH-SY5Y cells. Briefly, 1.0 × 106 SH-SY5Y cells in 100 μL resuspension buffer were electroporated with 12.0 μg shRNA plasmids into 150 × 20 mm tissue culture dishes containing 15 mL Opti-MEM medium containing 5% FBS. After optimization, transfection of SY-SY5Y cells were performed in triplicate for each shRNA plasmid and incubated for 72 hours.

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Manufacturer protocol

Download the product protocol from Qiagen for Attractene Transfection Reagent below.

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