ExoSAP Reaction

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The ExoSAP reaction "removes" the dNTP's and PCR primers from the PCR product to prepare it for Sanger Sequencing. It contains Exonuclease I and Shrimp Alkaline Phosphatase that chop up single stranded primers and inactivate dNTP's so they do not interfere with the sequencing reaction. The mixture is:

10 μl PCR product
3 μl ExoSAP (which is 1 μl of Exonuclease I and 2 μl of Shrimp Alkaline Phosphatase)

The mixture is heated to 37 C for 15 min then 80 C for 15 min. This is under the program "EXSAP" on the thermocycler.

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