果梅基因组DNA提取方法的比较及ISSR分析

以果梅品种鸳鸯梅、皇后梅和肖山选的幼嫩叶片为试材,提取果梅的DNA.针对果梅组织细胞体内含有较多的酚类、糖类及萜类等次生代谢物质的特点,采用传统的CTAB法、改良的CTAB法和改良的SDS法提取果梅基因组DNA,并比较其提取效果.结果表明:改良的CTAB法可获得高质量、高得率的基因组DNA,OD260/280比值在1.80左右.通过基因组DlNA-ISSR分析,完全满足试验要求.并进一步对改良的CTAB法的关键步骤作出具体的分析讨论.     

微生物发酵床猪舍空气微生物种群结构的研究

为了研究微生物发酵床猪舍空气微生物的组成,采用自然沉降法对猪舍空气微生物进行收集,利用16S rRNA序列分析及形态观察确定微生物的分类,揭示猪舍空气微生物的多样性。从猪舍空气中共分离到细菌60余株,经过16S rRNA基因序列的同源性比对,最终确定27个代表菌株进行后续分析。分析结果表明,27株细菌中包含芽孢杆菌属14个种（51.9%）,假单胞菌5个种（18.5%）,葡萄球菌4个种（14.8%）,苍白杆菌属、类芽孢杆菌属、赖氨酸芽孢杆菌属及单胞菌属各1个种;27株细菌中包含5种条件致病菌,6种有机物降解菌。采用SPSS分析将猪舍空气微生物按采集地点明显聚成6类：仅分布于猪舍下风处的菌、仅分布于猪舍上风处的菌、仅分布于猪舍外的菌、猪舍上风和下风处共有菌、猪舍下风处和猪舍外共有菌及3处共有菌。对猪舍空气微生物空间分布的研究显示,猪舍下风处微生物种类较多,有20种,多于猪舍上风处和猪舍外部,其中15种为下风处特有种。由于微生物以气溶胶的形式存在,猪舍内相邻位置具有相似的微生物组成。以上研究结果表明,微生物发酵床猪舍空气微生物种类丰富,且微生物分布与风向相关。     

几种内生真菌DNA提取方法的比较

通过3种内生真菌DNA提取方法的比较,即CTAB法、SDS法、改良的CTAB法,结果表明三种方法都能得到目的DNA,而且得率都较好,但改良的CTAB法效果最好,不仅DNA纯度高且质量也好.用改良的CTAB法所提取的DNA作为模板,进行PCR扩增,得到了清晰的、单一的扩增普带,完全可用于酶切、测序等后续实验.     

利用无机膜分离技术降低大米饮料沉淀率的研究

大米饮料是目前国内饮料厂家密切关注和开发的产品。大米饮料因采用富含淀粉质的大米作为原料,经水解液化后制备而成,货架期内极易发生沉淀分离现象,严重降低了产品的商品价值。采用无机膜分离技术处理大米饮料,选择合适处理工艺及参数,可除去大米饮料中微米级颗粒。研究结果表明,无机膜处理能有效解决大米饮料的沉淀问题,工艺技术稳定可靠,可取代离心分离等净化技术。     

Application of Chitosan to Technology of Clarification for Chinese Traditional Medicine Extraction

As the product of natural macromolecule chitin deacetylating, Chitosan is linear macromolecule carbohydrate and is widely applied to medicine, chemical industry and environmental protection, etc.. The distribution, general properties and clarifying principle of chitosan, and the application of chitosan to the clarification of Chinese traditional medicine extraction are described . Compared with ethanol sedimentation process, the chitosan clarification is a safe, of innocuity, cheap and fine clarifying process. Chitosan clarification process can replace the conventional ethanol sedimentation process as the method of fining Chinese traditional medicine extraction and preparation. The feasibility and the problem are discussed. And the advice for future researching orientation is put forward.     

三种DNA提取方法对检测草莓镶脉病毒稳定性的研究

草莓成熟叶片中含有大量的多糖、蛋白质、多酚和色素等物质,影响高质量DNA的提取和下游PCR反应的进行。为确定适用于PCR技术检测草莓镶脉病毒(Strawberry vein banding virus,SVBV)的DNA提取方法,从草莓叶片中提取DNA后,对样品中的SVBV进行特异性的PCR检测,比较确定不同DNA提取方法对PCR检测SVBV稳定性的影响。以感染SVBV的草莓叶片为材料,分别采用CTAB试剂盒法、高盐低pH法、改良SDS法提取草莓叶片的总DNA,将不同方法提取的DNA通过紫外分光光度法和琼脂糖凝胶电泳法进行测定,对DNA产量和质量进行分析。结果表明,3种方法均能获得较为完整的DNA片段,用CTAB试剂盒提取的DNA质量最高,改良SDS法提取的DNA质量较低。将不同方法提取的DNA经过PCR反应特异性检测SVBV后,将与目的片段大小一致的PCR扩增产物经序列测定及分析,证明与SVBV的序列相吻合,证明了方法的可靠性,其质量能够满足PCR特异性检测需要。     

Effect of Two Pretreatment Techniques on Preventing Membrane Fouling

Ultrafiltration membrane was used to filter the Yangtze River water by powder activated carbon (PAC) pretreated, such as Tai Lake water by coagulation-sedimentation pretreatment. The changes of organic polar content were compared in each process. Combined with the scanning electron microscope(SEM) photographs of the membrane, the mechanism of slowing down membrane fouling by PAC and coagulation-sedimentation pretreatment was studied. The results showed that hydrophobic organic compounds were the main factors of membrane fouling, while hydrophilic organics had less impact. When the dosing quantity of PAC filtered by ultrafiltration membrane was 20 mg/L to pre-treat the water samples of Yangtze River, and that of polyaluminium chloride was 25 mg/L for the pretreatment of Tai Lake water samples, the flux could be recovered effectively after backwashing compared with filtering the two raw water directly. The cake layer formed on the surface of the membrane was looser after pretreatment, and could be discharged easily by periodic backwashing, The cake layer could prevent hydrophobic organic matters deposit on the surface of membrane directly, which will slow down the membrane fouling and improve the chemical safety of the membrane effluent.     

棉花基因组DNA快速提取方法改良

选择2种简化的棉花基因组DNA提取方法并进行适当精简改良后与传统CTAB提取法进行实验对比的结果显示,改良SDS-CTAB法所提DNA得率较高,纯度好,但其操作步骤相对复杂。改良CTAB法操作更加简捷,但DNA得率相对较低,所得DNA溶液浑浊度较高。相对于传统的CTAB提取法,两种改良方法都能够更加快捷的得到足量且纯净的基因组DNA,完全能够满足转基因后代PCR筛选检测的要求。     

2种提取椰子树RNA方法的比较研究

椰子树组织中富含多糖和酚类化合物，为了获取高质量的RNA，笔者以椰子树叶片和根部为材料，比较分析CTAB法和SDS法提取RNA的效果。结果表明：CTAB法提取椰子树组织的总RNA效果较好，能有效克服酚类物质和多糖对RNA提取的影响。经OD值检测和电泳检测，总RNA的28 S和18 S条带清晰可见，纯度较高，完整性好，无明显降解，以此RNA为模板进行RT-PCR反应，能获得特异条带，表明该RNA完全可以用于后续的分子生物学操作。