# Knight:Reconstituting primers

(Difference between revisions)
 Revision as of 11:34, 14 March 2007 (view source) (→Notes)← Previous diff Current revision (10:12, 7 May 2010) (view source) Line 12: Line 12: ===Long version=== ===Long version=== - $Y\ \mu L\ =\ \frac{1}{25\ \mu L}*X\ nmoles\ primer\ =\ \frac{1*10^6\ \mu M}{25000\ nmoles}*X\ nmoles\ primer\ =\ 40\ *\ X\ nmoles\ primer$ + $Y\ \mu L\ =\ \frac{1 L}{25\ \mu moles}*X\ nmoles\ primer\ =\ \frac{1*10^6\ \mu L}{25000\ nmoles}*X\ nmoles\ primer\ =\ 40\ *\ X\ nmoles\ primer$ ==Notes== ==Notes==

## Procedure

Invitrogen recommends the following reconstitution procedure -

1. Centrifuge the tube for a few seconds to get all the DNA to the bottom of the tube.
2. To make a 25 μM stock, add YμL of sterile H2O to X nmoles of dry primer stock. (See equations below).
3. Allow to sit for 2 mins, then vortex for 15 secs.

### Short version

$Y\ \mu L\ =\ 40\ *\ X\ nmoles\ primer$

The number of nmoles of material in the tube (X) should be listed on the pages accompanying your primer order.

### Long version

$Y\ \mu L\ =\ \frac{1 L}{25\ \mu moles}*X\ nmoles\ primer\ =\ \frac{1*10^6\ \mu L}{25000\ nmoles}*X\ nmoles\ primer\ =\ 40\ *\ X\ nmoles\ primer$

## Notes

See Reconstituting primers for a lot of useful information including why you should use TE buffer rather than water for reconstituting primers.