Reaction of Biotin-BMCC with Sulfhydryl-Modified DNA

Reaction of Biotin-BMCC with Sulfhydryl-Modified DNA 

Since maleimide groups are highly specific for coupling to thiols in the pH range of 6.5 to 7.5, side reaction products can be avoided. The reaction is complete within two hours at room temperature. 

Prepare 10-20 pg of a sulfhydryl-containing oligonucleotide in 200 pi of 50 mM sodium phosphate, lOmM EDTA, pH 7.2 (the methods outlined in Section 2.2 of this chapter can be used to form the thiol group). 

Dissolve biotin-BMCC in DMSO at a concentration of 5 mg/ml. Prepare fresh. 

Isolate the biotinylated probe by ethanol/salt precipitation as described in Section 1 for nick translation (this chapter). 

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