We perform Sanger sequencing for varied range of PCR products. We usually employ ExoSAP for PCR product clean-up reaction. Due to unavoidable reasons we are looking for ExoSAP alternative.
Does ExoCIP from NEB perform equally good?
Technically what is the difference in the enzymatic activity of Calf Intestinal Alkaline Phosphatase and Shrimp Alkaline Phosphatase? Which enzyme has optimum performance for PCR product clean-up activity?
I think that both enzymes (CIP and shrimp AP) will work but the shrimp AP is less heat stable so inactivation of the shrimp enzyme will happen at a lower temperature and more quickly which is an advantage before moving on to the next step and more likely to maintain the integrity of the dna if you can use lower temperatures.
Paul Rutland Thanks for your message. I will better stick to Shrimp alkaline phosphatase.
I've had better results with the shrimp phosphatase as well. CIP is harder to deactivate and I had to add an additional clean-up step to get rid of the enzyme. Lost a lot of PCR product in the clean-up.
My vote is for classic ExoSAP-it reagent or shrimp AP as well.
Katie A S Burnette Thanks for sharing your experience with us. I would better stick to ExoSAP-IT.
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