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exosap_reaction [2019/10/15 16:30] (current)
floyd created
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 +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