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primer walking

A method for sequencing long (>1 kb) cloned pieces of DNA. The initial sequencing reaction reveals the sequence of the first few hundred nucleotides of the cloned DNA. On the basis of these data, a primer containing about 20 nucleotides and complementary to a sequence near the end of sequenced DNA is synthesized, and is then used for sequencing the next few hundred nucleotides of the cloned DNA. This procedure is repeated until the complete nucleotide sequence of the cloned DNA is determined.

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