10 Questions
What is the primary function of bacterial alkaline phosphatase (BAP) in gene manipulation?
BAP hydrolyzes 3’ and 5’ phosphate from nucleic acid (DNA/RNA), removing phosphate groups before end labeling and from vectors prior to insert ligation.
Why is calf intestinal alkaline phosphatase (CIP) used in gene cloning experiments?
CIP is used to remove phosphate groups from the 5’ end of DNA and RNA, preventing self-ligation and recircularization of vectors.
What is the effect of inorganic phosphates on bacterial alkaline phosphatase (BAP) activity?
Inorganic phosphates reduce BAP activity.
What is the characteristic that makes shrimp alkaline phosphatase (SAP) suitable for use in cloning?
SAP is highly specific and heat-labile.
At what temperature does bacterial alkaline phosphatase (BAP) show optimal activity?
65°C
What is the purpose of dephosphorylation in cloning experiments?
Dephosphorylation prevents self-ligation and recircularization of vectors, improving the efficiency of vector constructs containing desired inserts.
What type of enzyme is alkaline phosphatase?
Phosphomonoester
What is the source of calf intestinal alkaline phosphatase (CIP)?
Calf intestine
Why is it necessary to remove phosphate groups from vectors prior to insert ligation?
To prevent self-ligation and recircularization of vectors.
What is the characteristic that makes bacterial alkaline phosphatase (BAP) suitable for use in gene manipulation?
BAP is a thermostable enzyme that can hydrolyze 3’ and 5’ phosphate from nucleic acid (DNA/RNA).
Learn about the SAP enzyme, its properties, and applications in molecular biology, including DNA and RNA dephosphorylation, cloning experiments, and PCR product preparation.
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