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What are the requirements for plasmid DNA used for sequencing?
General requirements for sequencing plasmid DNA:
(1) High DNA purity (A260/A280 ratio 1.6–2.0). It must not contain mixed templates, RNA, genomic DNA, or proteins.
(2) Dissolved in ddH₂O. The solution must not contain impurities such as salts or chelators like EDTA, as these will interfere with the sequencing reaction.
How to purify PCR products?
The PCR product must first be run on an Agarose gel. The target band is excised and then purified using a gel extraction kit. The eluted product should be dissolved in ddH₂O.