ON-TARGETplus Human TET1 siRNA

siRNA / miRNA gene silencing Human - A172 TET1

Experiment
siRNA / miRNA gene silencing Human - A172 TET1
Product
ON-TARGETplus Human TET1 siRNA from Dharmacon
Manufacturer
Dharmacon

Protocol tips

Upstream tips
Seed 1 × 10^5 cells per well.
Protocol tips
Incubate transfection mixture for 10-15 min at room temperature.

Replace medium after five hours and incubate for additional 48 h in in 0.2% hypoxic conditions.

Publication protocol

Cultured A172 cells were plated in six‐well plates (1 × 105 cells per well), 1 day before the transfection so that they are around 80% confluent before transfection. Next day, the cells were transfected using Lipofectamine 3000 (Invitrogen) according to the manufacturer's instructions. Briefly, in different sterile microcentrifuge tubes, 100 nM siRNA against Tet1 (On‐target plus TET1 siRNA smart pool ID 80312, Dharmacon Inc, Colorado, United States, http://dharmacon.gelifesciences.com), Tet3 (Ambion, Invitrogen; ID for siTet3: 47240), or Control siRNA (On‐target plus non‐targeting siRNA, Dharmacon) were diluted in 1× opti‐MEM (Gibco). Lipofectamine 3000 (2 µl) was added to each and the mixture was immediately vortexed. This was incubated at room temperature for 10–15 minutes and media were added to the tubes to make up the volume till 1 ml. During this time the plates were washed twice with Dulbecco's phosphate‐buffered saline to remove the cell debris. After 15 minutes, the Lipofectamine 3000/siRNA/Opti‐MEM mix was vortexed and gently added over the plated cells. The plate was incubated at 37°C for 5 hours. After 5 hour, the media were changed and cells were further incubated for 48 hours in 0.2% hypoxic conditions and harvested for RNA and lysate preparation subsequently.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Human - A172 TET1 using ON-TARGETplus Human TET1 siRNA from Dharmacon.

Paper title
Hypoxia-Mediated Epigenetic Regulation of Stemness in Brain Tumor Cells.
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Manufacturer protocol

Download the product protocol from Dharmacon for ON-TARGETplus Human TET1 siRNA below.

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