Product Catalog

Use this 3´-phosphatase-free preparation of T4 polynucleotide kinase to:

• phosphorylate the 5´-ends of RNA or DNA 
• phosphorylate the 3´-ends of RNA to prevent cyclization
• add a 5´[³² P]-terminal label to 3’-CMP, forming 5´[³² P]pCp
• label 5´-hydroxyl ends of DNA with [γ-³²P]-ATP (either by direct phosphorylation or by phosphate exchange)
• phosphorylate the 5´-hydroxyl ends of oligonucleotides obtained from automated synthesizers
• phosphorylate the 5´-hydroxyl ends of linkers 
  

Brief description: T4 polynucleotide kinase catalyzes the transfer of the terminal phosphate group of ATP to the 5´-hydroxyl terminus of DNA or RNA. It also can catalyze exchange of 5´-terminal phosphate groups. However, in contrast to the normal  E. coli enzyme, this phage derivative lacks a 3´-phosphatase activity.

Source: phage T4-infected E. coli

Preventing cyclization of RNA
Addition of 3’-phosphate prevents cyclization and concatamerization of the RNA.

5´[³² P]pCp
This labeled nucleotide can serve as a substrate for 3’-end labeling of RNA with T4 RNA ligase.  

Phosphorylation of oligonucleotides
Oligonucleotides which are obtained from automated synthesizers lack a 5´-phosphate group, so they cannot be ligated to other polynucleotides. T4 polynucleotide kinase can add the necessary 5´ end, so the oligonucleotides can be ligated.

Phosphorylation of linkers
Linkers must be 5´-phosphorylated before they can be used in ligation reactions.

Solution, 10 U/μl, in buffer, pH 7.5 (25^oC)

• Polynucleotide Kinase;
  supplied with 10x reaction buffer
• Alkaline Phosphatase, shrimp;
  
• Rapid DNA Ligation Kit;
  
• T4 RNA Ligase;
  
• Klenow Enzyme;
  labeling grade, DNA Polymerase I, large fragment
• Terminal Transferase;
  recombinant
• Agarose Gel DNA Extraction Kit;
  For the elution of DNA fragments from agarose gels
• High Pure PCR Product Purification Kit;
  For purification of PCR reaction products