# Calculation help: pmoles 4MU synthesized per seedling Watch

1. Hi, I could do with some help converting my data from the fluorimeter used in assay for GUS activity into the form"pmoles 4MU synthesized per seedling". Calculations aren't my strong point, and I'm always paranoid about what I have done

Background:

1 fluroesence unit = 1nM
Sample tube volume used in fluorimeter: 1.5ml (1.4ml STOP buffer, 0.1ml from the reaction tube)
Reaction tube volume: 0.6ml (0.57ml Gembug, 0.03ml from the extract tube)
Extract tube: 10 seedlings ground into 1ml geb

Calulations:

Example fluorimeter reading: 84.92

First I must determine the concentration of 4MU (nM) in the cuvette from my reading, which I think is just the same thing since 1 fluroesence unit=1nM, so 4MU conc = 84.92nM.

Next using the 1.5ml volume used in the fluorimeter I have to work out the amount of 4MU present as pmoles of 4MU present.
Using the equation: moles=molarityxvolume (L)
I did: 84.92nm*0.0015L = 0.12738 nmoles of 4MU
To get in terms of pmoles *1000 = 127.38 pmoles of 4MU

Could you please check to see if what I've done so far is right? I have no faith in my math ability If I have gone wrong, could you please tell me how/why please and thank you.

Next I have to work how much of this is produced per seedling, which I have no idea how to do. It might be really obvious, but I feel like I've been staring at it so much, I just can't work it out anymore :')

I imagine it involves me doing something with the reaction volume and seedling number (I was thinking maybe (0.6*0.03)/10? and then maybe dividing the pmoles of 4MU by this number?) but I'm really not sure and figured I'd ask before I base the next part on a set of values that were wrong

Thanks again (And sorry if I was silly :') Don't be afraid to tell me its all wrong!)
2. Even if you can't help me with the full thing, helping me with any part of this is welcome :') Just telling me if what I've done so far is right would be helpful too
Thank you

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Updated: April 7, 2013
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