反应详情

The ten oligodeoxynucleotides shown in FIG. 3A were synthesized by the phosphotriester method and purified. 500 pmole of each oligodeoxynucleotide except 1 and 10 were phosphorylated individually in 20 μl containing 60 mM Tris-HCl, pH 8, 15 mM DTT, 10 mM MgCl2, 20 μCi of [γ-32P]-ATP and 20 units of polynucleotide kinase (P/L Biochemicals) for 30 min. at 37° C. This was followed by the addition of 10 μl containing 60 mM Tris-HCl, pH 8, 15 mM DTT, 10 mM MgCl2, 1.5 mM ATP and 20 additional units of polynucleotide kinase followed by another 30 min incubation at 37° C. Following incubation the samples were incubated at 100° C. for 5 min. 500 pmole of oligodeoxynucleotides 1 and 10 were diluted to 30 μl in the above buffer without ATP.

1. custompurified
2. additionThis was followed by the addition of 10 μl
3. customat 37° C
4. waitwere incubated at 100° C. for 5 min