I am new in the field of siRNA -and so on this forum- since my main field is Pharmaceutical Technology.
My research interest recently focused on the possibility of preparing a lipidic or polymeric carrier to ensure the stability of siRNAs and their transfection in neuronal cells, with the possibility of a future in-vivo use of such carriers.
I have some questions about handling of siRNA:
1) I read in this forum and in various protocols many different suggestions about how to treat glassware, surfaces and plasticware: since I don't have the possibility of baking my glassware and I already have a lot of non-sterile plasticware, do you think is effective to soak all these items in DEPC-water overnight, then autoclave everything for few hours to eliminate the DEPC? for how long do you think I can use these items if I store them in sealed plastic bags?
2) do you think that RNAse can be present in chemicals? I will use phospolipids, cholesterol, peptides (from chemical synthesis), and Sephadex G 50 to run a chromatography column: do you suggest to treat these items in some way before putting in contact with the siRNA?
3) syringes for human use can be considered RNAse-free (since they are sterile)?
4) do you suggest to work under a laminar flow hood or it is enough to pay attention to air currents, people around, using gloves and deeply cleaning every surface?
I know that you already answered to some of these questions for RNA extraction from cells, but I would like to know if conditions have be the same in my case, since the siRNA duplex (19 nt base pairs + 2 dT overhangs) with phosphorothioate modifications should be more stable against rnases.
Thanks for your attention and for your kindness,