Alkaline phosphatase and polynucleotide kinase for DNA and RNA dephosphorylation or phosphorylation.

FastAP Thermosensitive Alkaline Phosphatase T4 Polynucleotide Kinase (T4 PNK)
Applications
  • Dephosphorylation of DNA, RNA and oligonucleotides
  • Dephosphorylation of vector DNA
  • Dephosphorylation of proteins
  • Phosphorylation of PCR products
  • Phosphorylation of PCR primers and oligonucleotides
  • Labeling of sequencing primers
  • Detection of DNA modification by the [32P]-postlabeling assay
  • Removal of 3’-phosphate group
Substrate
  • 5’- and 3’-phosphorylated DNA/RNA
  • Oligonucleotides
  • dNTPs/NTPs
  • Phosphorylated proteins
  • ATP*
  • DNA/RNA
  • Oligonucleotides
  • 3’-NMP
  • 3’-phosphorylated DNA/RNA
  • 3’-P-oligonucleotides
  • Deoxynucleoside 3’-monophosphates
  • Deoxynucleoside 3’-diphosphates
Reaction catalyzed
  • Release of phosphate group
  • Transfer of the γ-phosphate from ATP* to the 5’-OH group of DNA/RNA, oligonucleotides, 3’-NMP
  • Exchange of phosphate group between 5’-P-oligo-/polynucleotides and ATP (in the presence of ADP)
  • Release of 3’-phosphate group
Reaction products
  • 5’, 3’-OH ends of DNA/RNA
  • Oligonucleotides
  • Deoxynucleosides / nucleosides
  • Dephosphorylated proteins
  • 5’-phosphorylated DNA/RNA
  • 5’-P-oligonucleotides
  • 3’, 5’-NDP
  • 3’-dephosphorylated DNA/RNA oligonucleotides
  • Deoxynucleosides
ss – single-stranded
ds – double-stranded
*Although ATP is routinely used as a phosphate donor, CTP, UTP, GTP, dATP, dTTP are equally effective.