s4215

siRNA / miRNA gene silencing Human - PANC-1 DNMT1/3b

Experiment
siRNA / miRNA gene silencing Human - PANC-1 DNMT1/3b
Product
s4215 from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
Seed 1 × 10^5 cells per well
Protocol tips
The DNMT1 siRNA sequences were 5′-GGAUGAGAAGAGACGUAGAtt-3′ and 5′-UCUACGUCUCUUCUCAUCCtg-3′, and the DNMT3B siRNA sequences were 5′-GCUCGUCUCCUAUCGAAAAtt-3′ and 5′-UUUUCGAUAGGAGACGAGCtt-3

Final concentrations - 7 nM

Add diluted siRNA to diluted Lipofectamine Reagent  

Incubate for 5 min at RT and add to cells. 

Replace medium after 6h and re-incubate cells for 2 days at 37°C.

Publication protocol

The DNMT1 and 3b-specific siRNA duplexes were chosen from the GenBank sequences (accession #NM_001379.1 and #NM_175849.1, respectively) and synthesized by Ambion Inc. (Austin, TX, USA). The DNMT1 siRNA sequences were 5′-GGAUGAGAAGAGACGUAGAtt-3′ and 5′-UCUACGUCUCUUCUCAUCCtg-3′, and the DNMT3B siRNA sequences were 5′-GCUCGUCUCCUAUCGAAAAtt-3′ and 5′-UUUUCGAUAGGAGACGAGCtt-3′. Negative control siRNA was also purchased from Ambion Inc. (Catalog #4390843). Final concentrations were 7 nM. The OptiMEM (Gibco BRL Inc.) and Lipofectamine 2000 Transfection Kits (Invitrogen) were used for siRNA transfections. Cells were plated at 1 × 105 cells per well in culture dishes for overnight growth and transfected by siRNA on the following day. Six hours after transfection, the medium was removed and replaced with DMEM, and the cells were allowed to grow for 48h. Then, viability, apoptosis, cell cycle and gene expression were assayed.

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

Reviews

s4215 from Thermo Fisher Scientific has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Human - PANC-1 DNMT1/3b using s4215 from Thermo Fisher Scientific.

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

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for s4215 below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Human - PANC-1 DNMT1/3b using s4215 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.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms