Establishing siRNA Experiments - controls, sequences...recommendations (Oct/13/2008 )
hallo all,
i want to establish an in vivo siRNA Experiment. does anyone of you know which controls have to be included (GAPDH siRNA...) and how many siRNA sequences should be tested in parallel?
thanks for suggestions.
-moljul-
For in vivo experiment, you don't need to use positive controls (such as siRNA for gapdh), a siRNA with scrambled sequence is good enough. For your target gene, one siRNA that works great in vitro is enough.
-pcrman-
QUOTE (pcrman @ Oct 14 2008, 02:29 AM)
For in vivo experiment, you don't need to use positive controls (such as siRNA for gapdh), a siRNA with scrambled sequence is good enough. For your target gene, one siRNA that works great in vitro is enough.
is it ok when i test 3-4 sequences in vitro, and take then the one which silences best to continue with in vivo experiments?
-moljul-
QUOTE (moljul @ Oct 14 2008, 07:23 PM)
QUOTE (pcrman @ Oct 14 2008, 02:29 AM)
For in vivo experiment, you don't need to use positive controls (such as siRNA for gapdh), a siRNA with scrambled sequence is good enough. For your target gene, one siRNA that works great in vitro is enough.
is it ok when i test 3-4 sequences in vitro, and take then the one which silences best to continue with in vivo experiments?
for invitroexperiments you have to have one universal positive and one universal negative siRNA...other than that one housekeeping control , either GAPDH or beta actin....and the strategy you are using to use the best invitro molecule into in vivo systems is by the book the right way....but u sure can have transfection hazards and also other reactions per say...
-laila-