Knight:Reconstituting primers

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Contents

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}{25\ \mu L}*X\ nmoles\ primer\ =\ \frac{1*10^6\ \mu M}{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.

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