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exosap_reaction

This prepares a PCR product from a PCR reaction for Sanger Sequencing.

The ExoSAP reaction “removes” the dNTPs and PCR primers from the PCR product. 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.

The result is an “ExoSAP'd PCR product” for Sanger Sequencing.

exosap_reaction.txt · Last modified: 2019/10/15 16:30 by floyd