Product Introduction
Shrimp alkaline phosphatase catalyzes the dephosphorylation of 5 'and 3' phosphate monophosphates of DNA and RNA, and also hydrolyzes ribose and deoxyribonucleoside triphosphate (NTP and dNTP). This phosphatase is widely used in molecular biology research, such as the removal of phosphate groups at the ends of DNA and RNA, for subsequent cloning and probe end labeling. In cloning, the linear vector can be dephosphorylated to prevent self-linking. Shrimp alkaline phosphatase can be completely and irreversibly deactivated at 65℃for 5 minutes, so it is not necessary to remove the thermal phosphatase after linking or end labeling. Based on these properties, the enzyme is an excellent alternative to the traditional dephosphorylase, calf intestinal alkaline phosphatase (CIP).
Application
1. Dephosphorylation of DNA and RNA
2. Prevent self-linking of cloning vector
3. Prepare 5′ end label template
4. Remove dNTP and pyrophosphate from PCR products
Precautions
(1) 1×SAPBuffer: 50mmbis-TrishclPH 6.0, 1mM
MgCl2, 0.1mMZnCl2, incubated at 25℃.
(2) Thermal inactivation: 65℃heating for 5 minutes, irreversible inactivation. This enzyme is commonly used
Dephosphorylation of enzyme cut vectors due to their irreversible heat inactivation properties. After the dephosphorylation reaction is completed, the product does not need to be purified again and can be directly used for subsequent linking reactions.
(3) Shrimp alkaline phosphatase can also be used to degrade free dNTP in PCR reactions for the preparation of sequencing templates and SNP analysis. It also requires no purification and can be used directly for downstream experiments.
Note:Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.