We describe here a method for the synthesis of oligonucleotides with block structure (padlock probes, primers for multiplex polymerase chain reaction (PCR), and ligation-independent cloning), based on the ligation of presynthesized parts by T4 DNA ligase. The advantages of this approach are: (i) suitability of the technology for any producer-from synthesis company to laboratory, (ii) high quality and adjustable scale of synthesis, and (iii) possibility of including any modified bases inexpensively in the common part of the oligonucleotide. Clear difference of sizes of products and substrates makes the synthesis amenable to automation. For large series of padlock probes, the price per one primer approaches the price of the locus-specific parts. We demonstrate the application of this method to two different tasks: preparative-scale production of padlock probes and small-scale synthesis of PCR primers.
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