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How to quantify oligonucleotide primers using nanodrop ND-1000 software


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#1 mercutio22

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Posted 09 September 2009 - 12:44 PM

Hello!

I need to verify if a vial containing DNA oligonucleotide primers has the same concentration as the labeled one.

Now, should I choose the regular DNA-50 option or ssDNA-33 option in order to do so?

I expect the molecules to be ssDNA, but it is conceivable that they fold into hairpins and form double helices right?

#2 mextorquens

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Posted 11 September 2009 - 03:40 AM

Hi!

In Nanodrop software, for the sample type, you can choose "Other". Select this one, and type 30 as the constant. You have to take in account that this number is an approximation. You can also calculate more precisely the concentration by using OligoCalc.

Hope I help you! :(

Melanie

#3 mercutio22

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Posted 11 September 2009 - 05:45 AM

Hi!

In Nanodrop software, for the sample type, you can choose "Other". Select this one, and type 30 as the constant. You have to take in account that this number is an approximation. You can also calculate more precisely the concentration by using OligoCalc.

Hope I help you! :(

Melanie

Melanie,

Thanks for your reply. I assume this constant is the molar absovity constant in Lambert and Beer's law relating Absorbance to molar concentration. The OligoCalc tool, doesn't seem to compute that. By the way, how is that value computed or determined empirically?

Sorry if thats too much. I am just curious.




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