primer concentration - (Oct/20/2006 )
Hi
Which is a useful primer concentration to use for a regular PCR?. I was using a slow amount, but reading some texts, I realised that I m using too low concentration in uM.
Could anyone help me?
It is suggested in most Polymerase datasheets to use 100-250uM. You can check on the datasheet of your polymerase or use something in this range
I use a final concentration of 0.2 uM in the PCR reaction, which for me works well for all my primers, but you may want to try a few concentrations to see what works best with your primers and PCR conditions. Using too high a concentration of primers can sometimes result in non-specific bands or a lot of primer dimers, so using a lower amount is a good idea.
I used different unit. I use in term of ng. Apparently, my lab is the weirdest one to use ng for primer concentration. =)
What are the difference between both?
I used around 300ng which showed primer dimers as well.
What are the difference between both?
I used around 300ng which showed primer dimers as well.
The amount of mol changes according to the number of nt. You could calculate the mol you are using: on the datasheet you receive with your primers it should be written how many moles they sent you and you should be therefore able to calculate the concentration of the primers you are using
Hi....
I use 100 uM stock primer concentration, and 100 to 250 uM seems to be high.
On the other hand, 0,2 uM is a slower amount of what I use.... I remember reading that around 5 uM is the recommended amount to start to set up a PCR rxn. Could anyone confirm that to me.
To the 100 to 250 uM, I ll check into the datasheets of my Polymerase.
Thanks and waiting for new recommendations
I use 100 uM stock primer concentration, and 100 to 250 uM seems to be high.
On the other hand, 0,2 uM is a slower amount of what I use.... I remember reading that around 5 uM is the recommended amount to start to set up a PCR rxn. Could anyone confirm that to me.
To the 100 to 250 uM, I ll check into the datasheets of my Polymerase.
Thanks and waiting for new recommendations
Typing mistake: it was nM, not uM. I'm very sorry!
Thank you for the verification of concentration.
into another website I ve found from 0'2 to 1 uM, but also the tip: less ys better!!!!!!
Thank you all!!!!!