5 replies to this topic

[uzhamd]

Hello all
I want to test enzyme activity at 100 and 105 C with total reaction mixture of 500 ul.
but cannot figure out how to perform it with out evaporation problem. can any one help.

Edited by uzhamd, 01 April 2012 - 09:14 AM.

[doxorubicin]

Can you use a PCR machine with a heated lid?

[hobglobin]

Or add a drop of mineral oil that covers the surface, such as in PCRs in machines without heated lid...the oils are still available (e.g. from Sigma Aldrich)
or perhaps just some Parafilm?

Edited by hobglobin, 01 April 2012 - 12:57 PM.

[phage434]

You may need to find a way to do the reaction under slight pressure, since the liquid will likely boil at those temperatures.  A screw capped vial immersed in heated oil would probably be pretty easy, but shield yourself from a potential rupture of the vial.

[uzhamd]

I tried in PCR but could not bcz it wont work on 100 & 105 C. But use of oils seems to be good option but would not it interfere with end product analysis in this case i use DNS method for estimation of reducing sugars (end production) immediately after the completion of incubation period.

[mdfenko]

you can use a hot block to get to 105C and beyond.