Abstract
Relative to conventional locked nucleic acids (LNAs), C5-functionalized LNAs with small entities at the C5-position display markedly higher (1) duplex thermostability with complementary single-stranded DNA/RNA targets, (2) target specificity, and (3) 3'-exonuclease stability. C5-Functionalized LNAs carrying a polarity-sensitive fluorophore enable sensitive and efficient discrimination of single nucleotide polymorphisms (SNPs) under non-stringent conditions. This unit describes protocols for chemical synthesis of an LNA uridine diol and corresponding C5-functionalized LNA uridine phosphoramidites. A procedure for incorporation of C5-functionalized LNA uridine phosphoramidites into oligodeoxyribonucleotides by automated DNA synthesis is also described.
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