摘要
本研究的目的是改进头孢菌素C产生菌顶头孢霉基因组DNA的提取方法。采用氯化苄法、液氮研磨法以及石英砂研磨法进行比较,从提取的DNA数量及纯度来分析,石英砂法要好于液氮研磨法,与氯化苄法效果相当,但避免了氯化苄的毒性。提取液的pH对结果影响不大,当pH为9.0～10.0之间时,收率稍高;EDTA浓度应大于40 mmol/L。经石英砂研磨破壁,获得了较高质量的染色体DNA,该法与液氮研磨法、氯化苄法相比廉价、快速、安全,且不影响其后续的分子生物学操作。
A new method in which quartzite is used to grind the mycelia to prepare the chromosomal DNA is studied. The solution（100 mmol/L Tris, 100 mmol/L NaCl, 40 mmol/L EDTA- Na2, 2% SDS,PH 9.0） as the extraction buffer, and quartz sand crashing the cell wall, can get a high qualitire chromosomal DNA. After analyzing the DNA sample extracted by homogenizing in liquid nitrogen and by Benzyl Chloride, it is found that when the quartzite grind method is adopted, the DNA is better whatever the amount and the purity is mentioned, and it is homology with the Benzyl Chloride, but avoided the toxicity of the Benzyl Chloride. It is also found that the pH of the extract liquid is not crucial to acquire the DNA, and when pH value is 9.0- 10.0, the DNA amout is big and the concentration of the EDTA must be higher than 40 mmoL/L. The DNA acquired with this method can satisfy the need of molecular analysis. Comparing with liquid nitrogen and benzyl chloricle method, this method is much cheaper, faster and safer.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2007年第5期8-11,共4页
Journal of Hebei Agricultural University
关键词
顶头孢霉
石英砂研磨
染色体DNA
提取
Cephalosporium acremonium
quartzite sand grind
chromosomal DNA
extraction
