TurboFect Transfection Reagents

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed ~5 × 10^4 cells	Dilute 1µg of DNA in 100µL of serum-free DMEM or other serum-free growth medium. Incubate 15-20 minutes at room temperature. Add transfection reagent/DNA mixture to samples and incubate at 37°C in a CO2 incubator.	Analyse expression after 24-48 hours

Upstream tips
Seed ~5 × 10^4 cells
Protocol tips
Dilute 1µg of DNA in 100µL of serum-free DMEM or other serum-free growth medium. Incubate 15-20 minutes at room temperature. Add transfection reagent/DNA mixture to samples and incubate at 37°C in a CO2 incubator.
Downstream tips
Analyse expression after 24-48 hours

Publication protocol

Small interfering RNA (siRNA) was performed to knock down M2AChR in H9c2 cells. The siRNA oligonucleotides were synthesized by GenePharma Co. Ltd (Shanghai, China). The siRNA sequence targeting M2AChR is as follows: 5'-GUGCUCAUCAAUACUUUCUTTAGAAAGUAUUGAUGAGCACTT-3'. A scrambled non-target siRNA was also used as a negative control. The detailed method was described previously [18]. Transfections were performed with 100 nM of each siRNA using TurboFect siRNA Transfection Reagent (Thermo Fisher Scientific, Cleveland, OH, USA) according to the manufacturer's instructions. Forty-eight hours after transfection, the efficiency of siRNA-mediated M2AChR knockdown was determined by Western blotting.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TurboFect Transfection Reagents below.

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