Can biotinylated oligonucleotides be phosphorylated with kinase?
Yes, 5' biotin labeled oligonucleotide can be phosphoyrlated with kinase. Teoule and coworkers have shown(1) that oligos labeled at the 5'-terminus are substrates for kinase. The oligos modified with reagents based on 1,2-diols are labeled to 50%, indicating that only one diastereomer is labeled, while those modified with 1,3-diol reagents are labeled to 100%. Bio-Synthesis can modified your oligo based on a 1,2-diol,e.g., Biotin TEG, DNP modifiers, or 1,3-diol, e.g., fluorescein, biotin modifiers. These reagent can be added several times at the 5'-terminus since they contain an alcohol group capable of further addition with other modifiers. Alcohol group in these modifiers can act as a substrate for T4 polynucleotide kinase for P32 labeling.
Reference:
(1) M.L. Fontanel, H. Bazin, and R. Teoule, Analytical Biochemistry, 1993, 214, 338-340.
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10/23/2013