Nucleic Acids and Oligonucleotides

Primers

Dissolve the forward and reverse external primers in H2O at a concentration of 100 pM (100 pmoles/pl). Dissolve the mutagenic primer in H2O at a concentration of 10 pM (10 pmoles/pl).

Template DNA

Superhelical, double-stranded plasmid DNA, purified by ethidium bromide-CsCl density gradient centrifugation (Chapter 1, Protocol 10 or Chapter 1, Protocol 11) or chromatography on a Qiagen resin (Chapter 1, Protocol 9). Dissolve the template DNA in TE (pH 8.0) at a concentration of 0.1 pg/ml.

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