| 鱼类血液基因组DNA提取方法优化 |
| Optimizing Method of Extracted Genomic DNA from Fish Blood |
| 投稿时间：2009-04-13 |
| DOI： |
| 中文关键词:鱼类 基因组DNA 提取 |
| 英文关键词:Fish Genomic DNA Extracting method Optimization |
| 基金项目:甘肃省自然科学基金(No.0803RJZA030)资助 |
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| 中文摘要: |
| 以冷水性虹鳟鱼抗凝冷冻全血为材料,分别采用氯仿-醋酸钾法、RGDE(Rapid genomic DNA extraction)法、改进的RGDE法、传统酚-氯仿法、试剂盒法提取基因组DNA,琼脂糖凝胶电泳检测显示改进的RGDE法可以获得较高质量的鱼类基因组DNA。进一步将改进的RGDE法提取血液基因组DNA的效果与传统酚-氯仿法提取血液、肌肉基因组DNA及试剂盒法提取血液基因组DNA的效果通过紫外分光光度计测定浓度和纯度进行比较。结果表明,改进RGDE法提取的基因组DNA平均浓度为1050ng/μL,极显著高于用试剂盒从血液中提取的DNA浓度,显著高于酚-氯仿法从血液和肌肉中提取的DNA浓度,DNA产量为1575μg/mL,A260/A280比值1.65~1.97。采用改进的RGDE法提取DNA,全过程只需2h,具有速度快、成本低、无污染、无需特殊设备的明显优点,而且随机扩增的结果稳定,电泳条带清晰。 |
| 英文摘要: |
| The genomic DNA extracted from frozen whole blood of rainbow trout with methods of chloroform-potassium acetate, rapid genomic DNA extraction （RGDE） , improved RGDE, traditional phenol-chloroform and DNA reagent kit respectively, and detected by agarose gel electrophoresis. The results showed that mothode of improved RGDE can obtain better genomie DNA of fish. Then, the effect of extracting with improved RGDE methods were compared by ultraviolet spectrophotometric method with other mothods such as traditional phenol-chloroform from blood or muscle and DNA reagent kit from blood. The results showed that the average concentration of DNA extracted with method improved RGDE was 1 050 ng/μL, and was very significant higher than that by DNA kit or by phenol- chloroform, and the average output of DNA was 1 575μg/mL, and the rate of A260/A280 was between 1.65 - 1.97. It only spent 2 hours to extract DNA with the method of improved RGDE, and there were many obvious merits such as quick speed, low cost, no pollution and no using specific installations and so on, and to amplify by random primers, the rusuhs was stable,and the electrophoresis banding was clear. |
| 刘哲,康鹏天,柴文琼,王太,刘晓敏.2009.鱼类血液基因组DNA提取方法优化[J].水生态学杂志,30(6):102-106. |
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