Resuspending Primers

Your primers will arrive as a lyophilized film at the bottom of a cryo-tube. To use them, you must resuspend them in autoclaved dH2O.

  1. Resuspend it to 100 uM/L concentration. For example, for an amount of 28.5 nm of lyophilized primer (the amount is provided by the vendor on the spec sheet and the side of the tube), add 285 ul water or TE buffer to get a 100 uM/L stock solution. Pipette up and down until the film dissolves (you can watch it through the bottom of the tube) or resuspend by vortexing.
  2. To prepare 10 uM/L (or 10 pm/ul) working solution, take 10 ul stock solution and add to 90 ul water.
  3. To use in a typical 50 uL PCR reaction, add 1 uL of working solution of each primer to the reaction. Adjust for reactions of different volume. 1 uL is also appropriate for a 12 uL sequencing reaction at the ICMB core.

-- Main.MichaelHammerling - 06 Jun 2011

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Contributors to this topic Edit topic MichaelHammerling, JeffreyBarrick, JuliePerreau, LindseyWolf, AurkoDasgupta
Topic revision: r4 - 2011-06-07 - 22:47:39 - Main.MichaelHammerling
 
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