Hs_PTEN_6 FlexiTube siRNA

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	CT116 colorectal carcinoma cells were sourced from the American Type Culture Collection. HCT116 cells were cultured in Roswell Park Memorial Institute 1640 (RPMI-1640) medium supplemented with 10% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc.), 50 U/ml penicillin/streptomycin and 2 g/l sodium bicarbonate. Transfection was done using Lipofectamin 2000. The siRNAs were designed to specifically target their corresponding transcripts. Designed with asymmetrical 5′base pair stabilities, these double-stranded oligos ensure entry of less tightly-bound antisense strand to the RNA-induced silencing complex while reducing risk of off-target effects by degrading the sense strand, which can be incorrectly processed by the RNA-induced silencing complex. The optimum concentration for each siRNA was 30 nM. RNA extraction with TRIzol (Invitrogen; Thermo Fisher Scientific, Inc.) of siRNA-trans-fected cells was performed according to the manufacturer's protocol, and the generated corresponding cDNA was used for RT-qPCR to validate the efficacy of the siRNAs.

Protocol tips

CT116 colorectal carcinoma cells were sourced from the American Type Culture Collection. HCT116 cells were cultured in Roswell Park Memorial Institute 1640 (RPMI-1640) medium supplemented with 10% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc.), 50 U/ml penicillin/streptomycin and 2 g/l sodium bicarbonate. Transfection was done using Lipofectamin 2000. The siRNAs were designed to specifically target their corresponding transcripts. Designed with asymmetrical 5′base pair stabilities, these double-stranded oligos ensure entry of less tightly-bound antisense strand to the RNA-induced silencing complex while reducing risk of off-target effects by degrading the sense strand, which can be incorrectly processed by the RNA-induced silencing complex. The optimum concentration for each siRNA was 30 nM. RNA extraction with TRIzol (Invitrogen; Thermo Fisher Scientific, Inc.) of siRNA-trans-fected cells was performed according to the manufacturer's protocol, and the generated corresponding cDNA was used for RT-qPCR to validate the efficacy of the siRNAs.

Publication protocol

CT116 colorectal carcinoma cells were sourced from the American Type Culture Collection. HCT116 cells were cultured in Roswell Park Memorial Institute 1640 (RPMI-1640) medium supplemented with 10% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific, Inc.), 50 U/ml penicillin/streptomycin and 2 g/l sodium bicarbonate. Transfection was done using Lipofectamin 2000. The siRNAs were designed to specifically target their corresponding transcripts. Designed with asymmetrical 5′base pair stabilities, these double-stranded oligos ensure entry of less tightly-bound antisense strand to the RNA-induced silencing complex while reducing risk of off-target effects by degrading the sense strand, which can be incorrectly processed by the RNA-induced silencing complex. The optimum concentration for each siRNA was 30 nM. RNA extraction with TRIzol (Invitrogen; Thermo Fisher Scientific, Inc.) of siRNA-trans-fected cells was performed according to the manufacturer's protocol, and the generated corresponding cDNA was used for RT-qPCR to validate the efficacy of the siRNAs.

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Manufacturer protocol

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