Phosphatases & Kinases

	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.