1994年
Efficient amplification of the HLA-DQA1 gene in single genomes using a semi-nested polymerase chain reaction
Japanese Journal of Legal Medicine
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- 巻
- 48
- 号
- 5
- 開始ページ
- 329
- 終了ページ
- 335
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
A semi-nested polymerase chain reaction (PCR) was introduced to amplification of the HLA-DQA1 gene, which is a single-copy gene, in a single genome. The limitation of template DNA for genotyping is 1 ng of genomic DNA, or more in the case of ordinary PCR. When reamplification with the same primers was performed, primer dimer was generated and the sensitivity was not improved. We designed a semi-nested primer for the second round of PCR. using the semi-nested PCR, more than 3 pg of template DNA could be amplified and typed. Furthermore, this method was applied to amplify DQA1 gene in single human sperm having haploid DNA, and followed by typing with sequence-specific oligonucleotide (SSO) probes. The semi-nested PCR technique was found to enhance the sensitivity of the amplification reaction and allowed the successful typing of the HLA-DQA1 gene. This is helpful for genotyping from samples with extremely small amounts of DNA, such as forensic or ancient DNA samples.
- リンク情報
- ID情報
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- ISSN : 0047-1887
- PubMed ID : 7807714
- SCOPUS ID : 0027940413