Latest revision as of 10:13, 21 July 2018

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

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 This prepares a PCR product from a [[PCR]] reaction for [[Sanger Sequencing]].
-The ExoSAP reaction "removes" the dNTP's 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:
+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]])