siRNA SKIL

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 30000 cells/well	The siRNA sequences (5′-to-3′) were: 1: GGCAAGUAAGUCCAUAUCATT (sense) and UGAUAUGGACUUGCCTC (antisense) 2: GGCUCACAGUAGUGGUAATT (sense) and UUACCACUACUGUGAGCCTT (antisense) siRNA concentration-50 nM Add INTERFERin® to siRNA duplexes and incubate for 10min at RT after homogenization.  Remove culture medium from cells and add incubate mixture.  Incubate the plate at 37°C for 24 to 72 h.

Upstream tips
Seed 30000 cells/well
Protocol tips
The siRNA sequences (5′-to-3′) were: 1: GGCAAGUAAGUCCAUAUCATT (sense) and UGAUAUGGACUUGCCTC (antisense) 2: GGCUCACAGUAGUGGUAATT (sense) and UUACCACUACUGUGAGCCTT (antisense) siRNA concentration-50 nM Add INTERFERin® to siRNA duplexes and incubate for 10min at RT after homogenization.  Remove culture medium from cells and add incubate mixture.  Incubate the plate at 37°C for 24 to 72 h.

Publication protocol

The knockdown of SKIL expression was done using small interfering RNA (AM16708, ID 107695) from Ambion (Ambion, Austin, TX, USA). The siRNA sequences (5′-to-3′) were:

1: GGCAAGUAAGUCCAUAUCATT (sense) and UGAUAUGGACUUGCCTC (antisense)

2: GGCUCACAGUAGUGGUAATT (sense) and UUACCACUACUGUGAGCCTT (antisense)

Silencer® Negative Control #1 siRNA (AM4611, Ambion, Austin, TX, USA) was used as a non-targeting control. Cells were seeded into 24-well plates (30000 cells/well) in four replicates and were transfected the following day with 50 nM SKIL siRNA and scrambled siRNA. INTERFERin TM (PolyPlus Transfection, Strasbourg, France) and Opti-MEM were used for cell transfection.

Growth curve analysis
Cells were plated on 12-well plate (30 000 cells on each well) as quadruplicates and each well was scanned daily using the Surveyor Software (Objective Imaging Ltd.) with a camera (Imaging Inc., Canada) attached to the Olympus IX71 (Olympus, Tokyo, Japan) microscope and the area of the attached cells in each well was computed using ImageJ Software (Wayne Rasband, National Institutes of Health, Bethesda, MD) and divided by the mean area for day 1.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Human - RWPE SKIL using siRNA SKIL from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for siRNA SKIL below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Human - RWPE SKIL using siRNA SKIL from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.