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Reagents & Solutions 800-356-9526 608-274-4330 enotes@promega.com
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FAQspeakShould I use Calf Intestinal Alkaline Phosphatase (CIAP) or Shrimp Alkaline Phosphatase (SAP) to dephosphorylate the 5′ ends of DNA?SAP is preferred for dephosphorylation of cloning vectors to prevent recircularization. SAP is easily heat-inactivated under the same conditions as many restriction enzymes and is more tolerant of non-optimal buffer conditions. SAP or CIAP can be used to dephosphorylate DNA before 5′-end labeling with T4 DNA Kinase, but SAP will be easier to inactivate. SAP is inactivated by heating at 65°C for 15 minutes. CIAP is inactivated by incubating at ≥56°C for 30 minutes in the presence of 5mM EDTA (pH 8.0), 0.5% SDS and 50µg/ml proteinase K. Alternatively, CIAP can be inactivated by incubating at 65°C for 1 hour or 75°C for 10 minutes in the presence of 5mM EDTA (pH 8.0). Products may be covered by pending or issued patents or may have certain limitations on use. Please visit our patent and trademark web page for more information. |
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