Here is my situation:
Instead of using synthetic double stranded mature microRNA from ambion, I am going to have both guide and passenger strands chemically synthesized. then I am going to use annealing buffer and annealing conditions from Metabion to get double stranded mature microrna. then i am going to use a transfection reagent( Effecten) to introduce microrna to cells.
1- has anyone been in the same situation before?
2- are micrornas prepared this way stable enough?
3- what chemical modifications can i use to increase the stability? would adding FAM/2'OM to 3' end change the stability or RISC involvement?
All the best