regarding primer designing

[priyatripuraneni]

hi all,

happy to be as a member of this station. i am here with some doubts regarding primer designing. please help me.

what is dG value given by the oligo analyser
what should be the optimum value of dG

thanks in advance
bye
priya

[said]

le protocole et les mecanismes

[moleculardude]

Hello there Priya!

and welcome to the forums:

dG is the free energy (G) value of the entire oligonucleotide.

I wouldn't worry about dG values between the two oligonucleotides.

[real time PCR]

Hi Priya,

the lesser the dG, bette it is. A low value will indicate that the bonds between the base pairs in oligos will break easily and won't interfere in the progress of a PCR reaction. Higher dG means you need to run PCR at a higher temperature

[real time PCR]

The dG calculations do not matter at higher Tm values

[real time PCR]

but specificity is one very important matter to be taken into account

[real time PCR]

And may be primer efficiency as well. You need to see if the template on which primers are being designed is OK and doen't folds upon itself. You know self complementarity