Nucleic Acids and Oligonucleotides

Forward primers (20 pM) in H2O

Reverse primers (20 pM) in H2O (20 pmoles/pl)

Template DNA

Long PCR works well on a variety of templates including recombinant PACs, BACs, cosmids, and bacteriophage  clones, as well as high-molecular-weight genomic DNAs. However, the quality of the DNA is paramount. The average length of the template DNAs (assayed by agarose gel or pulsed-field gel electrophoresis) should be at least three times greater than the length of the desired PCR product. The DNAs should also be extensively purified by equilibrium density centrifugation in CsCl gradients, followed by dialysis against TE (pH 8.0) (please see Chapter 1, Protocol 10).

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