不同保存方式下金龟甲虫DNA提取方法及28S rDNA序列分析

DNA Extraction and 28S rDNA Sequences Analysis of Scarab Beetle Specimens Preserved under Different Conditions

通过电泳检测和PCR扩增28SrDNA核基因序列的结果,比较了无水乙醇、75%乙醇、针插干制3种金龟甲虫标本的保存方式和SDS-蛋白酶K法、CTAB法、饱和NaCl法3种提取DNA方法.结果显示,SDS-蛋白酶K法、CTAB法提取3种保存方式的标本时,可成功提取DNA并有效扩增目的基因;饱和NaCl法仅能提取无水乙醇保存标本的DNA并扩增目的基因,而75%乙醇保存标本和干制标本的DNA提取和基因扩增均不理想.

The methods of DNA extraction and 28S ribosomal DNA sequences were compared from the specimens of scarab beetle Potosia（L.）brevitarsis（Lewis）which preserved in 100% ethanol,75% ethanol and kept as a dry-pinned specimen.The results show that the genomic DNA were obtained successfully and 28S rDNA sequences were amplified using polymerase chain reaction and sequenced using sequencing techniques from specimens preserved in three conditions by the extraction methods of SDS-protease K and CTAB.However,using the method of saturated NaCl,the specimens that have been fixed in 100% ethanol could yield successful genomic DNA extraction,amplification and sequence information of 28S rDNA sequences,yet preserved dry samples and 75% ethanol fixed samples were quite poor for DNA extraction and 28S rDNA sequences analysis.