为验证经高温加工过的鳖甲是否可以用DNA分子标记鉴定,选取经高温高压蒸煮过的中华鳖鳖甲和骨进行DNA提取,并用通用引物L1091和H1478扩增。结果表明经高温高压蒸煮过的鳖甲也可以提得较高质量的DNA,与从肌肉和药材鳖甲中提取的DNA比较没有明显差异,并可用作模板进行PCR扩增,得到约400bp基因片段;同时还讨论了DNA提取和扩增过程中应注意的问题,为分子遗传标记技术广泛应用于同类药材的鉴定积累了基础资料。

In order to ascertain whether autoclaved Trionyx sinensis Wiegmann could be authenticated by DNA molecular technology, DNA from boiled, steamed, and autoclaved shells and bones of the turtle were extracted and the template DNA amplified by universal primers L-1091 and H-1478. The results showed that there were no differences between the DNA obtained from the autoclaved material in comparison with those obtained from the boiled, steamed or traditionally processed crude drug. A 400 bp DNA fragment was amplified successfully from both boiled bone and muscles of crude drug under standard PCR conditions using the universal primers. The issue of DNA extraction and amplification were also discussed.

国家自然科学基金(No.3 9870913);江苏省教委科学基金(98KJB3 60010);江苏省“333工程”人才培养基金