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PCR product for sequencing


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#1 tashaD

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Posted 25 April 2010 - 05:09 AM

I have sent up several samples for sequencing. I am sending them away to be sequenced and they are also doing the pcr purification for me. I had a problem with fedex so my samples wont reach the sequencing company for at least another 5 days. The samples are currently in the freezer. Will this affect the quality of the sequencing? Should i prepare the samples again?

#2 ivanbio

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Posted 25 April 2010 - 07:52 AM

It is quite possible that if your samples are at room temperature while being shipped by FedEx that they will degrade before they reach the sequencing company. Wait until you get your results and see if it works, there is nothing you can do now. If when you get your results it did not work, then send them a new sample and this time make sure that the sample reaches them in 24 hours.

Alternatively to prevent this from happening again you can always dry your sample before shipping, that way even if it takes 5 days to reach the sequencing company the DNA will still be in good condition.

Ivan
Carlsbad, CA

#3 Micro

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Posted 26 April 2010 - 03:55 PM

Freezing the PCR product prior to sequencing is a bad idea. I work with high G+C content DNA and in the past have received unreadable mush for results when the PCR product is frozen.

Ivanbio idea is the best one... dried pellets of PCR product work best for transport.




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