荒漠植物PCR模板3种制备方法的初步研究

Three Methods of Preparing DNA Templates for PCR from Desert Plant

采用2×CTAB法、NaOH法及直接PCR扩增法,以准噶尔无叶豆〔Erem osparton songoricum（L itv.）Vass.〕为例,对沙生植物PCR模板制备方法进行了比较研究。结果表明：2×CTAB法提取的DNA产量及质量较高,PCR扩增谱带稳定、清晰,但方法繁琐,单个样品处理时间长,耗时耗力;NaOH法制备模板DNA简单快速、经济高效,在1-2 h可提取10-20个样品,比常规方法快3-5倍,并可产生稳定、可靠、重复性好的PCR扩增谱带;直接扩增法是一种更加快速、简便、廉价的制备方法,适宜制备大批量的PCR模板。研究结果对其他荒漠植物,尤其是叶片极端退化或珍稀濒危植物的PCR模板制备具有指导意义。

Three methods of preparing DNA templates for PCR of a desert plant, Eremosparton songorium （Litv.） for amplification, i.e. 2 × CTAB, NaOH and direct PCR amplification, are compared. The results show that the genome DNA obtained from three methods can satisfy the ISSR （ inter-simple sequence repeat） reaction. Among which, the high DNA quality and concentration can be obtained with 2 × CTAB method, and a clear PCR product can be gotten, but it costs more time and samples. Clear and reliable PCR bands can be produced with NaOH method although the DNA concentration is low, and the method is economical and effective. Normally 10 - 20 samples can be processed within 1 - 2 hours by applying this method, and the work efficiency can be increased by 3 - 5 times. Direct PCR amplification is the rapidest, simplest and lowest-cost method of extracting DNA, but the stability of PCR banding patterns is affected by the amount of plant material. Once the total amount of material is controlled and the PCR amplification system is developed, the Direct PCR amplification can be considered as the best method for a large-scale PCR test.