ExoSAP Reaction
From Genetics Wiki
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
What Links Here
- Methods (← links)
- Sanger Sequencing (← links)
