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Invitrogen™ Bacterial Alkaline Phosphatase

Catalog No. 18011015
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2500 U
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18011015 2500 U
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Catalog No. 18011015 Supplier Invitrogen™ Supplier No. 18011015
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Description

Includes

A vial of 10X dephosphorylation buffer [100mM Tris-HCl (pH 8.0)].

Removes 3' and 5' phosphates from DNA and RNA

BAP is active at 65°C for at least 1 hr. and is inactivated by phenol extraction

  • Source: Purified from E. coli C90
  • Performance and Quality Testing: Endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; dephosphorylation efficiency determined
  • Unit Definition: One unit hydrolyzes 1nmol of ATP in 30 min. at 37°C
  • Unit Reaction Conditions: 10mM Tris-HCl (pH 8.0), 1.5mM [γ-32P]ATP, and enzyme in 50μL for 30 min. at 37°C

Dephosphorylation of 5'-phosphorylated termini of vector DNA to prevent self-ligation; Dephosphorylation of 5' termini of nucleic acids prior to forward reaction with kinase; Cloning; Restriction Enzyme Cloning

Order Info

Shipping Conditions: Wet ice

Specifications

Content And Storage Bacterial Alkaline Phosphatase is supplied with a vial of 10X dephosphorylation buffer [100 mM Tris-HCl (pH 8.0)]. Store at -20°C.
Shipping Condition Wet Ice
Enzyme BAP
Quantity 2500 U
Product Type Alkaline Phosphatase

Frequently Asked Questions (FAQs)

Why are alkaline phosphatases used in cloning protocols? What is a typical protocol for dephosphorylation of nucleic acids?

Alkaline phosphatases are used to dephosphorylate the 5' ends of DNA. In cloning, it is used to prevent self-ligation of vector DNA. Standard ligation of DNA with ligase requires a 5' phosphate to be present on at least one of the ends being joined. When a DNA insert containing phosphates on both 5' termini is added to a dephosphorylated vector, the insert will be efficiently ligated into the vector, but the vector will not be able to self-ligate. Thus, dephosphorylation of vector lowers the number of background colonies containing vector without insert.

For Research Use Only. Not for use in diagnostic procedures.