I have siRNA-FAM labelled as -ve control. and I diluted it as company recommended to be 50uM (I use 1 ul equivalent to 50 picomole) ,and I want to make it 250 uM (so 1 ul would be equivalent to 250 picomole) instead.
so Can u suggest to me how I can make it more concentrated
I cannot increase the volume of used siRNA.
can I try to precipitate it again and reconstitute ?
Hi, you can ethanol precipitate your siRNA, reconstitute in a small volume of TDW, measure RNA concentration and adjust further to 250 μM with TDW. There would be some loss, though.
@Dooyoung Lee
Thanks for ur answer,
I want to increase conc to do electroporation experiment, and the use of ethanol ppt is not recommended by the company as ethanol might contain some impurities that will leak inside the cells due to electroporation and cause cell death.
Do u have other suggestion than ethanol ppt :)
You can try column-based kits for RNA concentration. I've used the ones from Zymo Research. Good luck :)
I have used vacuum centrifugation concentrator (MV-100 MicroVac Concentrator) to concentrate my sample and it worked fine for me
rotary evaporator, vacuum centrifuge, lyophilize, i´d recommend - on columns or by precip you may loose much of your probe
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