Difference between revisions of "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: | + | 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 | :10 μl PCR product | ||
:3 μl ExoSAP (which is 1 μl of Exonuclease I and 2 μl of Shrimp Alkaline Phosphatase) | :3 μl ExoSAP (which is 1 μl of Exonuclease I and 2 μl of Shrimp Alkaline Phosphatase) | ||
Revision as of 08:56, 17 July 2018
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.
What Links Here
- Methods (← links)
- Sanger Sequencing (← links)
