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Is it possible quantify dsDNA pre-sequencing with a kit in stepone plus qpcr

sequencing dsDNA qubit

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#1 Felippe Martins

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Posted 17 April 2014 - 12:38 PM

Hi everybody.

I have troubles in quantify dsDNA pre-sequencing in the Picodrop, so I heard about the Qubit, an equipament that quantify dsDNA by the fluorecence, for the use of this equipament we need a kit that has a fluorophore for DNA

I know that the StepONE Plus qpcr has a sensor of fluorescence, so does anybody know if its possible to quantify the dsDNA with this sensor using a kit with a fluorophore for dsDNA? my samples are product of a convencional PCR and to sequencing my samples I need the dna in 4ng/ul.

The Qubit in my country is a little expensive, so if i can use the stepone plus to quantify my dsDNA I will save some important money

Thanks for all


Edited by Felippe Martins, 17 April 2014 - 12:40 PM.


#2 mdfenko

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Posted 18 April 2014 - 04:04 AM

you can use one of these kits, from life technologies, with a microplate reader, for dna and these for rna.

 

qubit not required.


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#3 phage434

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Posted 18 April 2014 - 04:07 AM

I would guess that this would work fine. The major issue is not the sensor but the software for the qPCR machine, which may not easily report the observed fluorescence. Another alternative would be a plate reader with fluorescence detection. The picogreen reagent (Invitrogen/Molecular Probes) works well for DNA quantitation.







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