Patent Evaluation: Separation of biotinylated nucleotides from avidin-conjugated substrates using relatively mild conditions

Summary

Novelty: Pure single-stranded nucleic acid can be obtained by biotinylation of the target nucleic acid and recovery using an avidin capture system. This method has potential application in the fields of diagnostics, nucleic acid sequencing and forensics. It could also be used for the recovery of specific nucleic acid sequences from mixtures after chemical synthesis.

Chemistry: Nucleic acids can be biotinylated by a variety of procedures. For example, during chemical synthesis, a phthalimido triethyleneglycol phosphoramidite is incorporated at the 5′ end of the oligonucleotide and the phthalimide is converted into a primary amino group by hydrolysis. N-hydroxysuccinimidobiotin is then attached to the free amino group. The biotinylated nucleic acid is subsequently recovered by complexing with avidin. Release of the biotinylated nucleic acid from the biotin-avidin complex involves heating at above 60°C for several minutes. The released single-stranded nucleic acid can then be detected by a variety of methods including amplification, and use of complementary probes.