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water-droplets in my cycler !!! - experienced such thing ??? (Apr/26/2011 )

opening the cycler today morning, to bring out my first round PCR tubes,
realised there are water droplets on the heat-block ...
exactly on the edge-wells ...

so i was " ????? "
is it condensed water ??? But, if so ... what is the source ???

grateful to any insights ...

p.s : is this a sign that the cycler is getting out of date ?

-nightingale-

If this was a typical pcr protocol, it held the samples after cycling at 4C indefinitely. That was probably 10-12 hours. With any leakage of air into the sample holder, condensation of humidity will occur on the block. This is not surprising, nor worrying, but expected.

-phage434-

phage434 on Tue Apr 26 19:48:47 2011 said:


If this was a typical pcr protocol, it held the samples after cycling at 4C indefinitely. That was probably 10-12 hours. With any leakage of air into the sample holder, condensation of humidity will occur on the block. This is not surprising, nor worrying, but expected.


thank you :)
this was the first time i see such thing, so i was worried :S
thanks ...

-nightingale-

nightingale on Tue Apr 26 19:57:03 2011 said:


phage434 on Tue Apr 26 19:48:47 2011 said:


If this was a typical pcr protocol, it held the samples after cycling at 4C indefinitely. That was probably 10-12 hours. With any leakage of air into the sample holder, condensation of humidity will occur on the block. This is not surprising, nor worrying, but expected.


thank you :)
this was the first time i see such thing, so i was worried :S
thanks ...


Some manufacturers recommend holding at 10C. Less condensation. If the condensation seeps into the circuit boards, you'll have a dead machine. 10C is just as good as 4C for preserving your samples for short periods

-neuropath-

neuropath on Sun May 1 15:19:46 2011 said:


Some manufacturers recommend holding at 10C. Less condensation. If the condensation seeps into the circuit boards, you'll have a dead machine. 10C is just as good as 4C for preserving your samples for short periods

I think the main reason for this recomendation is to prolong the lifetime of Peltier blocks, which get exhausted by maintaining low temperatures for longer time.

-Trof-

thanks alot for the valuable info. given :)
i will keep an eye on the cycler.

neuropath: & what does "short period" means, overnight for example ???

-nightingale-

nightingale on Mon May 2 19:35:34 2011 said:


thanks alot for the valuable info. given :)
i will keep an eye on the cycler.

neuropath: & what does "short period" means, overnight for example ???


The longest I've tried is overnight around 12 hrs. I hardly need to do that these days after I discovered Kapa Biosystem's fast enzymes. At 1kb extension per sec (yes, sec!), your typical run is over in less than 30 min. I use their slower enzyme (cheaper) but its still 2kb/min. As for pricing, its comparable with other brands

P/S I do not work for Kapa or own their shares!

-neuropath-

The longest I've tried is overnight around 12 hrs. I hardly need to do that these days after I discovered Kapa Biosystem's fast enzymes. At 1kb extension per sec (yes, sec!), your typical run is over in less than 30 min. I use their slower enzyme (cheaper) but its still 2kb/min. As for pricing, its comparable with other brands

P/S I do not work for Kapa or own their shares!


per second ! ... that's really FAST !!!

p.s : are u sure of that ???!!! ;)

-nightingale-

I read the protocol, it says:

1 sec at 72 C for amplicons ≤1 kb, 15 sec/kb at 72 C for amplicons >1 kb

but then, the website also mention: Fragments up to 5 kb may be amplified from plasmid or lambda DNA with KAPA2G Fast, but fast amplification of genomic targets >3.5 kb is not recommended. (http://www.kapabiosystems.com/products/name/kapa2g-fast-pcr-kits)

@neuropath: I wonder how long is your fragment that you need 12 hours to do your PCR last time.

-adrian kohsf-

adrian kohsf on Thu May 5 05:43:17 2011 said:


I read the protocol, it says:

1 sec at 72 C for amplicons ≤1 kb, 15 sec/kb at 72 C for amplicons >1 kb

but then, the website also mention: Fragments up to 5 kb may be amplified from plasmid or lambda DNA with KAPA2G Fast, but fast amplification of genomic targets >3.5 kb is not recommended. (http://www.kapabiosystems.com/products/name/kapa2g-fast-pcr-kits)

@neuropath: I wonder how long is your fragment that you need 12 hours to do your PCR last time.


It wasn't the reaction time. It was the cold soak at the end of the run. That's how this topic started

-neuropath-