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question about bigdye


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4 replies to this topic

#1 claritylight

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Posted 20 February 2009 - 04:14 PM

How long can a sequencing reaction using BigDye be stored at 4 degrees Celsius? Can the reaction only be stored overnight at 4 degrees or will it be ok over a period of days at 4 degrees?

Edited by claritylight, 20 February 2009 - 04:25 PM.


#2 perneseblue

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Posted 20 February 2009 - 04:37 PM

you are better off just freezing it. Frozen it will keep for a year or so.
May your PCR products be long, your protocols short and your boss on holiday

#3 claritylight

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Posted 20 February 2009 - 04:48 PM

you are better off just freezing it. Frozen it will keep for a year or so.


The stock of bigdye is always frozen.

But when it is added into a sequencing reaction, can that be kept at 4 degrees for 24-48 hours without degradation or what is the timeframe? I ask because my sequencing reaction takes place on a thermocycler & the end of the reaction, it goes to 4 degrees Hold. Sometimes I can't always get to it to place on the sequencer right away, so I need to know how long I can keep it at Hold without going bad.

Edited by claritylight, 20 February 2009 - 04:50 PM.


#4 perneseblue

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Posted 21 February 2009 - 04:11 AM

I have kept the sequencing reaction at -20 C over the weekend without degradation. But I don't think I have kept the reaction at 4 C for any length of time.
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#5 phage434

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Posted 21 February 2009 - 07:59 AM

Stored samples need to be protected from light. I would guess (but have no experience) that keeping samples at 4C for a weekend would work, but -20 definitely works.




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