Chaisomchit Sumonta, Wichajarn Rattanawadee, Chowpreecha Sarunya, Chareonsiriwatana Wiyada
National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand.
Southeast Asian J Trop Med Public Health. 2003 Sep;34(3):641-5.
We have developed an efficient and simple method for extracting and purifying genomic DNA from dried blood stored on filter paper. The quality of the genomic DNA extracted is tested by PCR amplification of a 255-bp fragment of the PAX8 gene sequence and the PCR products are determined for further genetic studies by single strand conformation polymorphism (SSCP) analysis. Larger DNA sequences of the 674-bp of the PAX8 gene and the 1,039-bp of the human beta-globin gene, a housekeeping gene, have also been amplified from the extracted DNA, thus indicating the high quality of the genomic DNA extracted by the developed method for subsequent genetic studies of any gene of interest. The method developed can also be used for the purification of genomic DNA from dried blood specimens stored under different conditions. Moreover, the genomic DNA products can be stored for long-term use due to the highly purified procedure. Therefore, the method is efficient and appropriate for the extraction and purification of genomic DNA from dried blood specimens, which has become an increasingly important tool for genetic and epidemiological studies.
我们已经开发出一种高效且简单的方法,用于从存储在滤纸上的干血中提取和纯化基因组DNA。通过对PAX8基因序列的255bp片段进行PCR扩增来测试提取的基因组DNA的质量,并通过单链构象多态性(SSCP)分析确定PCR产物,以进行进一步的基因研究。还从提取的DNA中扩增了PAX8基因的674bp和管家基因人β-珠蛋白基因的1039bp的更大DNA序列,从而表明通过所开发的方法提取的基因组DNA质量很高,可用于后续对任何感兴趣基因的基因研究。所开发的方法还可用于从存储在不同条件下的干血标本中纯化基因组DNA。此外,由于纯化过程高度精细,基因组DNA产物可长期保存使用。因此,该方法对于从干血标本中提取和纯化基因组DNA是高效且合适的,这已成为基因和流行病学研究中越来越重要的工具。