| Abstract: | Haplotypes of the β-like globin gene cluster in individuals with sickle cell disease appear to be important as prognostic factors for the severity of the disease. The Hpa I polymorphic site 3′ of the β-globin gene is very often involved in haplotyping. However, this restriction site cannot be ascertained by conventional PCR, as it is in the middle of a long repetitive sequence. Long distance PCR was employed to amplify this Hpa I-containing region. By positioning either one or both primers outside the repetitive sequence, specific amplification was achieved. Moreover, sufficient amplification can be obtained by using a white cell lysate instead of purified DNA, thereby significantly reducing the amount of blood needed, and the total procedure time. |
