Bacterial Alkaline Phosphatase
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Bacterial Alkaline Phosphatase
Invitrogen™

Bacterial Alkaline Phosphatase

Bacterial Alkaline Phosphatase (BAP) removes 3´ and 5´ phosphates from DNA and RNA. BAP is active at 65°C for atRead more
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Catalog number 18011015
Price (USD)
366.00
Each
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Price (USD)
366.00
Each
Add to cart
Bacterial Alkaline Phosphatase (BAP) removes 3´ and 5´ phosphates from DNA and RNA. BAP is active at 65°C for at least 1 h and is inactivated by phenol extraction.

Applications:
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.

Source:
Purified from E. coli C90.

Performance and Quality Testing:
Endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; dephosphorylation
efficiency determined.

Unit Definition:
One unit hydrolyzes 1 nmol of ATP in 30 min. at 37°C.

Unit Reaction Conditions:
10 mM Tris-HCl (pH 8.0), 1.5 mM [γ-32P]ATP, and enzyme in 50 μl for 30 min. at 37°C.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Shipping ConditionWet Ice
EnzymeBAP
Quantity2500 U
Product TypeAlkaline Phosphatase
Unit SizeEach
Contents & Storage
Bacterial Alkaline Phosphatase is supplied with a vial of 10X dephosphorylation buffer [100 mM Tris-HCl (pH 8.0)]. Store at -20°C.

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.