ON-TARGETplus Human SMU1 (55234) siRNA - Individual

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	siRNA concentration-200 nM. Incubate the diluted oligonucleotide with diluted Oligofectamine mixture for 15-20 minutes at room temperature. Mix well and incubate the cells at 37°C in a CO2 incubator for 48 hours and add growth medium containing 3X the normal concentration of serum without removing the transfection mixture. Incubate cells for 48-72 h at 37°C in a CO2 incubator

Protocol tips
siRNA concentration-200 nM. Incubate the diluted oligonucleotide with diluted Oligofectamine mixture for 15-20 minutes at room temperature. Mix well and incubate the cells at 37°C in a CO2 incubator for 48 hours and add growth medium containing 3X the normal concentration of serum without removing the transfection mixture. Incubate cells for 48-72 h at 37°C in a CO2 incubator

Publication protocol

Control siRNA and pre validated SMU1 siRNA were purchased from Dharmacon (Catalogue no: J-021129-10). TRC lentiviral SMU1 shRNA (RHS4533-EG55234), DDB1 shRNA (RHS4533-EG1642), CUL7 shRNA (RHS4533-EG9820) and RNF40 shRNA (RHS4533-EG9810) were purchased from Dharmacon. Transfection was performed twice, 24 h apart, with 200 nM siRNA using Oligofectamine reagent in accordance with the manufacturer's protocol (Invitrogen). shRNAs were transfected transiently using PEI (Invitrogen) in BOSC23 packaging cells along with packaging vectors. 48 h post transfection, the viral medium was collected and added to the target cells along with polybrene (8 mg/ml). 48 or 72 h post infection, cells were collected and processed for various assays and immunoblotting was performed with the specific antibodies to check the efficiency of knockdown.

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Manufacturer protocol

Download the product protocol from Dharmacon for ON-TARGETplus Human SMU1 (55234) siRNA - Individual below.

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