Light Cycler

[Robert Hartley]

In article <39EEB969.9D0B35A6 at med.monash.edu.au>, Paul Cullen
<Paul.Cullen at med.monash.edu.au> wrote:


Hello Paul.
> Why does glass in the capillaries stuff up PCR using standard buffers?

You need BSA in the mix to prevent enzyme denaturation on the borosilicate
capiliaries. I don't have a light cycler but do have something similar IE
a Rapidcycler.  The borosilicate will also absorb Mg2+ so you need a conc
of min 3mM. 4mM should be OK for a standard reaction.

I should say that I have no afiliation with the folowin company BUT:::::
DNamp.com do a nice enzyme mix (I don't think they can call it a light
cycler mix) that has a range of premix concs with BSA. I'm sure Duncan
will be alng in a sec. :-)
> 
> We had a light cycler on loan and I found that I could not even
> reproduce many reactions that I carry out routinely?

see above
> 
> Another interesting question is why does it take more cycles?

new to me?
> 
> Just curious!

Cheers
Bob; Sunny Scotland (very) :-)
PS I'm back with the forcasts after breaking my news again. :-)

-- 
Robert Hartley, Centre for Cell Engineering,Joseph Black Building
University of Glasgow, Glasgow G12 8QQ Tel: ++44 (0)141 330 4756, 
Fax: 0141 330 3730 mailto:rh at mblab.gla.ac.uk
Web : http://www.gla.ac.uk/Inter/CellEngineering