2-mercaptoethanol

Posted: Mon Aug 04, 2008 6:33 pm

I am thinking of using 2-mercaptoethanol with a few specimens that have shown no peak (even wiping out standard!). I am wondering what concentrations have people used successfully in the past? I am thinking of adding it to the standard Galbraith's buffer recipe... perhaps along with PVP 40 (which I am already using).

Thanks for any advice,
Alaa

Posted: Thu Aug 07, 2008 1:34 pm

Hi Alaa,

In the LB01 buffer it is used at 15 mM. However many other researchers have used it at different concentrations. I suggest that you go through the FLOWer database (http://flower.web.ua.pt/) for further info.

You may also consider using 1% PVP-10 together with the mercaptoethanol. Other antioxidants as metabisulfite or DTT can also be used instead of the smelly mercaptoethanol.

Best,

JoÃ£o

Posted: Thu Aug 07, 2008 1:52 pm

Thanks for the tips!

Next week I am trying 10 mM sodium metabisulfate in Galbraiths for several species that have showed peaks, but where the standard peaks shifted dramatically. This is because for some other species I have found that using woody plant buffer has reduced that shift. Unfortunately, I've also frequently been getting very low event numbers with WPB (if I increase material I get too much debris and wide cvs). So, I thought adding the metabisulfate to Galbraith's might give me the best of both worlds... just a guess. I have a lot of tinkering to do.

(As an aside, I have found that for some woody genera, for instance Berberis, woody plant buffer indeed dramatically improves results over Galbraith's+PVP. Hooray!)

I am reserving the heavy (smelly) artillery for the species that only have debris in either buffer. I will try the merc. at 15 mM, with 10% PVP, as you suggest.

Thanks again!

Alaa

Posted: Thu Aug 07, 2008 2:16 pm

I am glad that you are using different approaches for different material. Also, it is nice that for some species the WPB works well. Smile

I look forward to hear from you and I hope that the analyses that you are going to perform next week are good. I also hope that you are able to put these results out soon as I look forward to read about your work.

All the best,

JoÃ£o