Dear Dr Donahue, dear Dr Phatak,
Hope your day is going well?
I' am writing to you because I' am interested in the pull down experiment that you conducted in your article (oncogene, 2016) and specifically the design of the miRNA mimics for these experiments.
From the protocol that you published "Biotinylated micro-RNA pull down assay for identifying miRNA targets" you mention Dharmacon for the synthesis of the mimics. Did you order the mimic duplex directly from them?
You also refer to the original protocol from Ulf Andersson (2007) and I was just wondering if you followed the tips from the article such as Phosphorylation of the 5' of each strand, having a "UU" overhang, mutation on the guide strand to destabilise it etc... when you designed the mimics for your experiments ?
I would like to thank you in advance for the help,
Best wishes,
Geoffray
10/16/2018 9:34:31 PM Reply