Pre-designed siRNAs - Using Dharmacon's and Ambions guranteed siRNAs (Jun/27/2005 )
I am trying to figure out whether its worthwhile to purchase the pre-designed siRNAs from Dharmacon or Ambion. Has anyone have any experience with either one? And does the inhibition as great as they say?
I trying to target K-ras, H-ras, and N-ras and as from the Ambion's pre-designed siRNAs, all of them target outside the mutation hot spots in Codon 12, 13, and 61. Is there a problem with targeting these spots or close to these areas?
You may want to make sure what pre-designed siRNA really means. Pre-designed could mean that siRNAs are designed before you order it (not verified), or siRNAs are experimentally verified. If it is the latter, then order it. For every gene, it is advised to have two siRNAs for transfection.
It is not a good idea to target a mutation hot spot unless you know the type of mutation in a certain cell, because any mismatch between the siRNA and its target potentially affects its function depending on where the mismatch occurs.
Hi. I just wanted to warn you about the G3PDH positive control that Ambion uses with their pSilencer kit -- it doesn't work! I went round and round with them, was repeatedly told that it was a transfection problem, but they eventually said that it just doesn't work in rat and mouse cells. I wasted 6 months of my life on this before I went to Dharmacon -- their new G3PDH knock-down construct knocks down 99%. Its wonderful! I wish that I had gone with Dharmacon in the first place.
thank you BECKYB for warning. are you talking about Ambion's Silencer GAPDH siRNA template set(cat no#4800) supplied with the silencer siRNA construction kit? please reply, i am ready to use this one for synthesising GAPDH siRNA to be used in rat cardiomyocytes, so plz...........