pSilencer 2.1-U6 Hygro

Protocol tips

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	SH-SY5Y, Dulbecco’s modified Eagle’s medium (DMEM, 12100-046; Gibco-BRL) supplemented with 10% heat-inactivated fetal bovine serum (FBS, SH30088.03; Thermo Scientific HyClone), 100 U/ml penicillin, 100 µg/ ml streptomycin and 2 mM glutamine. Cells were maintained in a humidified 10% CO2 atmosphere at 37°C. A human Beclin 1 shRNA hairpin (target sequence: CTC AGG AGA GGA GCC ATT T) was cloned into the HindIII and BamH1 sites of pSilencer 2.1-U6 Hygro. Beclin 1 shRNA and control scrambled shRNA plasmids were transfected into SH-SY5Y cells and selected with 50 µg/ml Hygromycin.

Protocol tips

SH-SY5Y, Dulbecco’s modified Eagle’s medium (DMEM, 12100-046; Gibco-BRL) supplemented with 10% heat-inactivated fetal bovine serum (FBS, SH30088.03; Thermo Scientific HyClone), 100 U/ml penicillin, 100 µg/ ml streptomycin and 2 mM glutamine. Cells were maintained in a humidified 10% CO2 atmosphere at 37°C. A human Beclin 1 shRNA hairpin (target sequence: CTC AGG AGA GGA GCC ATT T) was cloned into the HindIII and BamH1 sites of pSilencer 2.1-U6 Hygro. Beclin 1 shRNA and control scrambled shRNA plasmids were transfected into SH-SY5Y cells and selected with 50 µg/ml Hygromycin.

Publication protocol

SH-SY5Y, Dulbecco’s modified Eagle’s medium (DMEM, 12100-046; Gibco-BRL) supplemented with 10% heat-inactivated fetal bovine serum (FBS, SH30088.03; Thermo Scientific HyClone), 100 U/ml penicillin, 100 µg/ ml streptomycin and 2 mM glutamine. Cells were maintained in a humidified 10% CO2 atmosphere at 37°C. A human Beclin 1 shRNA hairpin (target sequence: CTC AGG AGA GGA GCC ATT T) was cloned into the HindIII and BamH1 sites of pSilencer 2.1-U6 Hygro. Beclin 1 shRNA and control scrambled shRNA plasmids were transfected into SH-SY5Y cells and selected with 50 µg/ml Hygromycin.

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Download the product protocol from Thermo Fisher Scientific for pSilencer 2.1-U6 Hygro below.

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