RNA extraction from low amount of human tissue - Trizol-silica columns (Feb/08/2008 )
I have to perform a RNA extraction from samples of 1-5mg of decidua (endometrium). Which method do you recommend?
Someone tell me that I can do it with Trizol method with glycogen, but I thought that it would be better to make it with silica columns like qiagen (RNeasy Micro Kit) or MN (NucleoSpin® RNA XS).
Which are the pros and cons of using this methods?
Thanks a lot
trizol with glycogen ensure to get all the RNA species both big and small ones. Moreover, using glycogen helps to pellet the RNA.
Using a column : you can get quicker big rnas (i mean >100-120ntd) but you'll losse part of it in the flow through of the column.
So if you choose the second option, i would hardly suggest you to do an RNA extraction on the flow through with the use of glycogen too. The advantage is that you have a separation of small rnas from the total rna. Moreover during quantitation, you are not so poisonned by all the rRNAs.
Finally if you want to do siRNA analysis, you have an enrichment of small RNAs species.
so in first instance i would go for trizol+glycogen.
If you want to focus on a particular RNA, then go for silica kit and choose the appropriate fraction for your analysis.