Assessing Pre-miR transfection efficiency - Without a fluorescent microscope (Mar/03/2010 )
Hi,
I'm currently performing Pre-miR (Ambion) transfections, but I have a bit of a problem: We do not have a fluorescent microscope so we cannot measure transfection efficiency using the labelled Pre-miRs. In stead I have been using RT-PCR to measure the increase in expression of the miRNAs in question and have been getting some pretty decent results.
This measure does not seem to be terribly quantifiable in terms of percentage transfection efficiency, does anybody have any ideas or examples on how to convert this to something publishable?
Baars01 on Mar 3 2010, 11:25 PM said:
I'm currently performing Pre-miR (Ambion) transfections, but I have a bit of a problem: We do not have a fluorescent microscope so we cannot measure transfection efficiency using the labelled Pre-miRs. In stead I have been using RT-PCR to measure the increase in expression of the miRNAs in question and have been getting some pretty decent results.
This measure does not seem to be terribly quantifiable in terms of percentage transfection efficiency, does anybody have any ideas or examples on how to convert this to something publishable?
you can try FCM. rt-pcr and wb also ok .
You don't need to show data on transfection efficiency in your paper. There is absolutely no need. If you are working with cell lines, the transfection efficiency can easily reach 100%.