PCR without thermal cycler? - (Nov/22/2009 )

Hi everyone!

I'm new here, just joined and already have questions

Well, I am a grad.student and I am trying to know how could I do a PCR within 24 hours and no thermal cycler?

is there a way?

thanks a lot for your help!

Pia on Nov 22 2009, 02:44 PM said:

Actually you only need three heat sources with annealing, elongation and denaturation temperatures, a clock and then put your cups manually from one source to the next for the needed time...water bathes or heat blocks are possible as heat sources....
if I remember right the first PCRs were done this way. If you do it don't forget the oil drop on the reaction mix to prevent evaporation and condensation at the lids...

i second on what hob says and i have observed it works better in a heating block (dry heat ) than a wet water bath.
But this is a very very tedious job. I wonder why you would like to do it!!!
Best luck anyways!!

I would think you could make your life much easier by bribing the lab down the hall with cookies, and borrowing their thermal cycler?

also, does your institution have a surplus department with old equipment?

good luck

Hmmm, nice! But why?

Have a look at this, i think

http://www.nature.com/nature/journal/v227/...df/227027a0.pdf

Chimp,
Have you noticed that article is almost 40 years old???
Things have changed.
Stagnation is not a virtue!!!

aimikins on Nov 25 2009, 03:31 AM said:

I quite like the bribe idea...cos I done it before. Unfortunately I bought Pizza, share them some boxes filtered tips...just to get to use 2 hours of that machine for 3 days.

I'd recommend talking to your supervisor about either getting a cycler for your lab or formalizing an agreement with the lab that has one. The single greatest expense in a lab is the workers' time, and manually performing PCR with water baths/heating blocks is an unbelievably inefficient practice. If you're just doing standard PCR, the cyclers aren't that expensive. Also nave a look at Global Medical Instruments or Lab-X for used/reconditioned equipment if your lab is on a tight budget.

I too would do the bribing thing! U asked for doing 1 PCRin 24 hours and you can actually do a weeks work in a day if u manage to convince some lab which has a PCR! Think practically...if you want to have some result which is reproducible, try to go with a machine!!

Or if the point is to prove that u can do a PCR without a cycler, then go ahead and show that u can do the work the cycler does in an hour if u work for a day

gogreen on Dec 1 2009, 09:50 AM said:

Or try the ebay thing, we bought once a 3x thermocycler from 1996 at ebay for about 100 euros, without heading lids but three blocks in one machine. It still works...