9 replies to this topic

[Pia]

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!

[hobglobin]

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

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!

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...

Edited by hobglobin, 22 November 2009 - 06:02 AM.

[Prep!]

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!!

[aimikins]

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

[Chimp]

Hmmm, nice! But why?

Have a look at this, i think

http://www.nature.co...df/227027a0.pdf

[molgen]

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

[Adrian K]

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

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

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.

[gfischer]

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.

[gogreen]

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

[hobglobin]

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

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

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...