一种高效提取虎耳草科植物基因组DNA的方法

An High-effective Method of Extracting Genomic DNA from Saxifragaceae Plants

[目的]探索从虎耳草科植物中提取DNA的有效方法。[方法]采用改进的CTAB法,从11种虎耳草科植物中提取DNA。以提取的DNA为模板,利用通用引物"psbAF"和"trnHR"对虎耳草科植物叶绿体DNApsbA-trnH片段进行PCR扩增。[结果]通过该方法提取的DNA纯度较高,质量较好。用所得DNA进行psbA-trnH扩增的产量高,可用于后续的测序等分析。对山地虎耳草的PCR产物纯化后进行测序,得到262 bp的序列。将其与GenBank中的虎耳草属其他植物的psbA-trnH序列进行比对分析,证实该序列为目标psbA-trnH片段的区域。[结论]该方法可有效去除次生物质对DNA的干扰,提取的基因组DNA可用于叶绿体psbA-trnH测序分析和其他遗传学分析。

[ Objective] The research aimed to explore the effective method of extracting DNA from Saxifragaceae plants. [ Method] DNA was extracted from 11 species of Saxifragaceae plants by using the modified CATB method. With the extracted DNA as template, the universal primers ＂psbAF＂ and ＂trnHR＂ were used to make PCR amplification on psbA-trnH fragment in chloroplast DNA from Saxifragaceae plants. [Result] DNA with higher purity and better quality was obtained by this method. The yield of PCR amplification on psbA-trnH with the obtained DNA was high and it could be used in analysis such as the subsequent sequencing. The PCR products of Saxifraga mentana H. Smith were purified and sequenced, and a sequence with the size of 262 bp was obtained. It was compared with psbA-trnH sequence in other plants Saxifraga L.on GenBank website, which proved that this sequence was target regiou of psbA-trn H sequence. [ Conclusion] This method could wipe off the disturbance of secondary substances to DNA and the extracted genomic DNA could be used in sequencing analysis of chloroplast psbA-trnH and other genetic analysis.