ExoSAP Reaction
From Genetics Wiki
The ExoSAP reaction "removes" the dNTP's and PCR primers from the PCR product to prepare it for sequencing. It contains Exonuclease I and Shrimp Alkaline Phosphatase that remove single stranded primers and inactivate dNTP's. 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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- Sanger Sequencing (← links)