primer walking

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Food Biotechnology

A method for sequencing long (>1 kbp) cloned pieces of DNA. The initial sequencing reaction reveals the sequence of the first few hundred nucleotides of the cloned DNA. Using this, a new primer of about 20 nucleotides is synthesized, which is complementary to a sequence near the end of sequenced DNA, and used to sequence the next few hundred nucleotides of the cloned DNA. This procedure is repeated until the complete nucleotide sequence of the cloned DNA is determined.

UN Food and Agriculture Organization — Glossary of Biotechnology for Food and Agriculture