三种全血基因组DNA提取方法的比较

目的 :比较三种不同的DNA提取方法提取人全血基因组DNA对DNA提取效率、纯度以及PCR扩增的效果影响。方法 :分别应用中山医科大学达安公司DNA提取液 ,珠海黑马医学仪器有限公司DNA提取液 ,胍盐酸法三种不同的方法提取人全血基因组DNA。结果 :三种方法提取的DNA纯度平均分别为 :1.4 8,1.5 1,1.5 6 ,各组间差异无显著性。 2 0 0 μL全血中所提取DNA总量平均分别为 1.6 μg ,2 .3μg ,4 .1μg。用 0 .8%琼脂糖凝胶电泳鉴定胍盐酸法制备的DNA效果较其它两种方法好 ,PCR扩增效果差异不明显。结论 :胍盐酸法提取的DNA总量明显高于其它两种方法 ,提取效率高 ,为三种全血基因组DNA提取方法中之首选。     

琼脂糖凝胶中DNA回收技术的探讨

从琼脂糖凝胶电泳分离的条带中回收DNA是常用的分子生物学技术。本试验通过比较常用的几种DNA片段的回收方法,发现用改进的离心法回收琼脂糖凝胶中的DNA片段效果较好,成本低,操作简单。并探讨了回收大片段DNA时,回收率较低的原因。     

PCR及电泳实验的单课时解决方案

“多聚酶链式反应扩增DNA片段”在高中生物新课程中占有重要的地位,但是PCR扩增和琼脂糖凝胶电泳的实验过程通常需要2h～3h才能完成,所以很难满足中学课时紧的要求,从而阻碍了在中学教学中的推广。针对以上问题,北京川布兰生物技术开发有限公司从研制相关试剂盒出发,将PCR扩增（21min）和琼脂糖凝胶电泳（5min-10min）控制在30min之内完成,从根本上解决了实验过程繁琐耗时长的问题。     

生化分析中常用的惰性载体——琼脂糖

琼脂糖作为优良的惰性载体和高分子筛在近代生物化学的定性、定量分析中,在对微生物蛋白质等生物材料的分离技术中得到广泛的应用。本文对琼脂糖在平板电流法、免疫扩散法、免疫电泳法、凝胶过滤法、亲合层洗法等生物技术中的应用,以及对如何在实验室中从琼脂中除去琼脂果胶制取琼脂糖做了论述。     

琼脂糖凝胶电泳装置及方法的改进

对现有的琼脂糖凝胶电泳装置进行了改进.以有机玻璃板等材料,制作出一套成本低廉、结构简便的琼脂糖凝胶电泳装置.经试用,具有操作简便、提高缓冲液的利用率、节约实验经费等优点,能够满足教学与科研的需要.     

Lipases in the castor bean seed of Chinese varieties：Activity comparison,purification and characterization

The lipase activities in the three main Chinese castor bean varieties [ZiBi 05 （ZB 5）,TongBi 06 （TB 6）,JinBi 02 （JB 2）] were investigated.Two lipases were found in extracts from the endosperms of all the three varieties.One lipase is located in the lipid bodies,showing higher activity at pH 4.5 and exhibiting the highest activity at day 1 of germination.The other lipase is located in the particulates,showing higher activity at pH 9.5 and particularly active at day 4 of germination.The lipid bodies from ZB 5 show the highest lipase activity at day 1 and pH 4.5 among the three varieties during the 8 d of germination.The acid lipase located in the lipid bodies of ZB 5 of germination 1 d was purified to homogeneity by DEAE-sepharose CL-6B and butyl-sepharose CL-4B chromatography after isolation,delipidation and solubilization,with a purification factor of 16.1 and overall activity recovery of 17.7%.The characterizations （such as optimum pH,pH stability,optimum temperature,thermal stability,effect of metal ions,organic solvents stability,substrate specificity） of the purified acid lipase were conducted in detail.A combination of the characterized properties may make this enzyme a useful biocatalyst for industrial application.     

Effect of agarose content on microstructures and mechanical properties of porous silicon nitride ceramics produced by gelcasting

<正>Porous Si3N4 self-reinforce ceramics were prepared by gelcasting using agarose solutions. By changing the agarose content in the slurries, the porous silicon nitride ceramics with different porosities, α→β-Si3N4 phase transformation, and mechanical properties were obtained. When the agarose content changed from 0.2% to 0.8% (w/w, based on powder), the porosities increased from 10.3% to 21.4%, while the fracture strength decreased from 455 to 316 MPa and the fracture toughness decreased from 6.6 to 5.5 MPa·m1/2. Many fibrous β-Si3N4 grains grown from the internal wall of the round pores is the typical microstructure of the gelcasting porous silicon nitride ceramic. Both elongated β-Si3N4 grains and suitable interfacial bonding strength contributes to high fracture toughness by favoring crack deflection and bridging. The growth mechanisms of fibrous grains resulted from the synergy of solution-diffusion-reprecipitation and vapor-liquid-solid (VLS).     

DNA片段的PCR扩增和琼脂糖凝胶电泳实验组织与安排

DNA扩增和DNA琼脂糖凝胶电泳实验首次推广到中学课程教学中。由于中学课时有限、班级人数多、试剂微量、试剂价格较高等诸多实际情况的制约,教学实验组织、指导和安排显得更为重要。在川布兰公司的大力支持下,2009年我校9个班近400个学生顺利完成了此项实验。每班40人共分8个小组,先做琼脂糖凝胶电泳实验(用上一个班所扩增的样品),再做DNA片段的PCR扩增实验,最后观察核酸条带。扩增前模板质粒由学生加入,在实验准备和安排上,尽量给每一个学生操作的机会。此次实验所用试剂盒为川布兰公司提供,总计费用878元。     

“酵母细胞的固定化”实验改进与探究

对“酵母细胞的固定化”实验中海藻酸钠的配制、酵母细胞的活化、发酵产物的检测等细节进行了优化。蠕动泵实现了凝胶珠制备的自动化、标准化,新型发酵装置实现了对发酵过程的定量、实时监测且显著缩短了实验周期。通过合作探究摸索出了琼脂糖替代海藻酸钠固定酵母细胞的方法,辩证地认识了固定化技术的优劣。