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What are the requirements for direct sequencing of PCR products?
(1) The amplification product must be specific, with a single band. If non-specific amplification products are present, it is generally difficult to obtain good sequencing results.
(2) Gel extraction and purification must be performed.
(3) DNA purity (A260/A280) should be between 1.6–2.0, with a concentration ≥ 50 ng/µL.