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.
EF0651,EF0652,EF0654,EK0031,EK0032
Ordering information
EF0651,EF0652,EF0654,EK0031,EK0032