Haplotypes of the human β-like globin gene cluster: Analyzing the 3′ Hpa I polymorphic site by long distance PCR
✍ Scribed by Sujit Sheth; Sergio Piomelli; Xiuhua Wang
- Publisher
- John Wiley and Sons
- Year
- 1995
- Tongue
- English
- Weight
- 293 KB
- Volume
- 50
- Category
- Article
- ISSN
- 0361-8609
No coin nor oath required. For personal study only.
✦ Synopsis
Haplotypes of the P-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 P-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.