Product Catalog
Alkaline Phosphatase 20 U/µl | |
| Catalog Number | Pack size | |
|---|---|---|
| 11097075001 | 1,000 U 20 U/µl, high concentration | Order and Price Information |
Application
Use this preparation of calf intestinal alkaline phosphatase to remove 5´-terminal phosphates from DNA or RNA. Removal of these phosphate groups is especially useful for enhancing cloning efficiency, since it prevents linearized vector DNA from self-ligating.
Note: This preparation has a high concentration of enzyme (20 U/μl), which is convenient for large-scale experiments. A lower concentration (1 U/μl) preparation is also available.
Product Description
Brief description: Alkaline phosphatase hydrolyzes 5´-monophosphate groups from both DNA and RNA. It can also hydrolyze 5´-diphosphate and 5´-triphosphate groups from RNA.
Source: calf intestine
Inactivation: Add 1/10 volume of EGTA to the reaction mix and incubate at 65oC for 10 min. To ensure complete inactivation of enzyme, extract the mix with phenol/chloroform/isoamyl alcohol to remove all protein.
Background Information
Preventing self-ligation of vectors during cloning
If a
linearized vector lacks a 5´-phosphate, it cannot ligate to itself or form
concatamers that contain multiple copies of the vector. Thus, the product of the
ligation reaction is predominantly recombinant DNA (vector + DNA insert), rather
than religated plasmid.
Contents
Solution, 20 U/μl, in buffer, pH 7.6 (20o C)
Additional Information
Product Literature
Instructions for Use and Material Safety Data Sheets
Associated Products
- Sma I from Serratia marcescens Sb;
- Rapid DNA Ligation Kit;
For rapid and efficient DNA ligation.
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