I am using MDA-MB-231 cell line in 6 well plate (300.000 cell/well) and I want to transfect them with 50nM and all the information I had is the mount is 5nmol.
I am using RNAiMax.
They gave you 5 nmoles. If you put this in 1 liter you will have a 5 nM solution. If you put it in 1 ml you get a 5micromolar solution. You probably want to have about a 10 to 20 micromolar stock solution. So, add 500 microliters to your 5nmoles of inhibitor and you will get a 10 micromolar solution.
this solution you have to dilute 1:200. let's say you have a 6 well plate, you probably use about 2 ml of medium. So, you need 10 microliters of your 10micromolar stock solution to prepare the RNAimax transfection mix. add this transfection mix to you cells that are cover with 2 ml of medium.
Your concentration in this medium will be 50nm inhibitor.
The only missing information is what volume of water/buffer was used to dissolve your miRNA.
They gave you 5 nmoles. If you put this in 1 liter you will have a 5 nM solution. If you put it in 1 ml you get a 5micromolar solution. You probably want to have about a 10 to 20 micromolar stock solution. So, add 500 microliters to your 5nmoles of inhibitor and you will get a 10 micromolar solution.
this solution you have to dilute 1:200. let's say you have a 6 well plate, you probably use about 2 ml of medium. So, you need 10 microliters of your 10micromolar stock solution to prepare the RNAimax transfection mix. add this transfection mix to you cells that are cover with 2 ml of medium.
Your concentration in this medium will be 50nm inhibitor.
Thanks Tomas, but again, the end concentration now is 10 micromolar in 500 microliter and this means 10.000 nanomolar in 500 microliter. So, if I need 50 nM it should be 25 microliter from the stock solution??
you need a 1:200 dilution to get from 10000 to 50 nM. if you have 2000 microliters of medium: 2000/200=10. 10 microliters. It is technically not 10000 nmoles in 500 microliters but 500 microliters of 10000nM solution.
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