一种从琼脂糖凝胶中回收DNA片段的简单电洗脱装置

我们设计了一种简单电洗脱装置,从琼脂糖胶中回收ＤＮＡ。该装置由两个带旋盖的小管、两块透析膜和一个凝胶屏障组成。在电场作用下,ＤＮＡ从凝胶中迁移出来,通过凝胶屏障进入由凝胶屏障和透析膜组成的回收小仓。用微量吸样器收集ＤＮＡ,乙醇沉淀和清洗。该法ＤＮＡ的回收率约８５％;回收的ＤＮＡ可用于基因工程常规实验。     

从琼脂糖电泳凝胶中回收DNA的几种简便方法

介绍两类从普通琼脂糖电泳凝胶中回收ＤＮＡ的简便、快捷、高效且廉价的方法．第一类为电泳洗脱法．方法ａ：利用１．５ｍＬ微量离心管、ｌｍＬ吸头、尼龙网膜和透析膜做成的一个小装置,快速有效回ＤＮＡ,最终回收率为７０％左右．方法ｂ：不用ＤＥＡＥ－纤维素膜,而用透析膜在凝胶中作出横隔挡在ＤＮＡ条带前,最终回收率为５０％左右;第二类为冰冻融解法,最终回收率也在５０％左右．如果联合使用冰冻融解法和电泳洗脱法,回收率可进一步提高至９０％．     

人琼脂糖电泳凝胶中回收DNA的几种简便方法

介绍两类从普通琼脂糖电泳凝胶中回收ＤＮＡ的简便,快捷,高效且廉价的方法,第一类为电泳洗脱法,方法ａ．利用１．５ｍＬ微量离心管,１ｍＬ吸头,尼龙网膜和透析膜做成的一个小装置,快速有效回ＤＮＡ,最终回收率为７０％左右,方法ｂ：不用ＤＥＡＥ－纤维素膜,而用透析膜在凝胶中作出横隔挡在ＤＮＡ条带前,最终回收率为５０％左右,第二类为冰冻融解法,最终回收率也在５０％左右,如果联合使用冰冻融解法和电泳洗脱法,回收     

PCR特异产物回收纯化方法的比较

方法：采用三种方法对苹果褪绿叶斑病毒RT-PCR的特异DNA产物进行回收纯化。目的：针对不同情况,选择适宜的回收纯化方法。结果：用普通琼脂糖替代低融点琼脂糖,回收纯化后产物的浓度及纯度与低融点琼脂糖法基本一致,完全可以用普通琼脂糖替代低融点琼脂糖进行DNA片段的回收纯化,从而降低成本,简化操作。玻璃奶法的回收纯度明显高于低融点琼脂糖法和普通琼脂糖法,且更快速安全,是采用普通琼脂糖法还是采用玻璃奶法回收纯化DAN片段应以实际需要而定。     

一种快速简单的质粒DNA纯化方法

本文介绍一种快速获取高纯度质粒DNA的方法。本法对煮沸提取法进行改良后,快速提取质粒DNA粗制品,然后用国产滤纸从琼脂糖凝胶中回收纯化质粒DNA。同时对本法所提取的质粒DNA的回收率、浓度、纯度及可能的用途进行验证和讨论,证明此法简易,所得样品纯度高,可直接用于转化、酶切和基因克隆。     

CTAB结合DNA凝胶回收试剂盒提取食用菌DNA

目的:为探索从食用菌子实体中快速分离和纯化DNA的方法.方法:以三种栽培的食用菌为材料,采用CTAB结合DNA凝胶回收试剂盒进行基因组DNA的分离和纯化.结果:与对照相比,结合法提取DNA样品的电泳条带更规则、清晰,并且DNA的酶切效率显著高于对照.结论:该结合法是一种快捷、高效提取纯化食用菌子实体DNA的方法.     

黄檗（Phellodendron amuranse）叶片总RNA提取方法研究

以黄檗(Phellodendron amuranse Rupr.)叶片为材料,分别利用改进的盐酸胍法、Trizol法、CTAB法提取黄檗叶片总RNA,通过RNA产率、纯度、电泳图谱等分析,确立了1种从黄檗叶片中快速分离总RNA的方法。研究结果表明,改进盐酸胍法所提取的总RNA的A₂₆₀/A₂₈₀为1.928,28S和18S条带清晰谱图完整性好,而且具有产率高、时间短、成本低的特点,所提取的总RNA适用于mRNA分离、cDNA文库的建立、Northern杂交等分析。     

山茶花叶片DNA提取及RAPD反应体系的研究

山茶花（Camellia japonica L.）是我国特产的名贵花卉之一,由于其品种繁多,在系统分类和品种鉴定上存在一定的难度。本研究针对山茶花的DNA提取方法和RAPD扩增体系进行了摸索,旨在为山茶花在DNA水平的分类鉴定和分子生物学方面研究打下一定的基础。实验采用改进的CTAB法提取山茶花叶片DNA,得到的DNA纯度较高,OD₂₆₀/OD₂₃₀值为1.90～2.0,OD₂₆₀/OD₂₈₀值为1.80～1.83,得率也较高,一般在150～200 μg·g^-1左右,片段完整性较好,大于20 kb,符合分子遗传实验的要求;在山茶花RAPD扩增条件的研究中,发现20 μL反应体系中加入100 ng DNA、125 μmol·L^-1 dNTP(each)、1.2 μmol·L^-1 Mg²⁺、1×Buffer、0.5 μmol·L^-1引物、1 U Taq酶最为合适,退火温度一般设为37℃左右,扩增产物的电泳条带数多、清晰、明亮。     

一种从琼脂糖凝胶上回收DNA的高效、快捷、经济的方法

目前在国内分子生物学实验室所采用的多种从琼脂糖凝胶上回收DNA的方法普遍存在着各种问题。本文介绍一种新的从琼脂糖凝胶上分离和提取DNA简易装置。实验表明用这种装置回收DNA具有高效、快捷和经济的优点,非常适合在国内实验室普及推广。     

狭叶坡垒基因组总DNA的提取和纯化方法研究

以狭叶坡垒叶片为材料,对基因组总DNA的提取和纯化方法进行了研究,并对改良CTAB法、高盐低pH法和改良SDS法进行比较.结果表明,改良的CTAB法更适合狭叶坡垒基因组总DNA的提取,且硅胶干燥30 d的叶样和新鲜叶样所得的DNA几乎没有区别.再对改良CTAB法的水浴时间进行探索,发现150 min是较为合适的水浴时间.对比酚纯化法和试剂盒纯化法,发现试剂盒纯化损失的DNA少,得率高,是一种简便、快速、安全的纯化方法.     

Simultaneous electroelution and concentration of DNA fragments from agarose gels

A rapid and inexpensive method for the electroelution of DNA fragments from agarose gels is described. DNA fragments were separated by agarose gel electrophoresis and visualized by staining with ethidium bromide. Selected DNA fragments were placed into electroeluter tubes capped with dialysis membrane and electroeluted into a small volume of buffer using a conventional horizontal gel electrophoresis apparatus. The method successfully eluted and concentrated DNA fragments with molecular weights ranging from 2.7 to 13.9 MDa in 3 h.     

简便实用的琼脂糖凝胶回收DNA片段方法

介绍一种简便实用的DNA片段回收方法,与以前所报道的DEAE-纤维素膜电泳法、透析袋电洗脱法、低融点琼脂糖凝胶法、凝胶冻融法等相比,所需器材简单、操作简便、回收率高、成本低。回收的DNA片段在进一步克隆和测序中表现出较好的效果,是一种适合于科研和教学的实验方法。     

Recovery of DNA from agarose gels using a modified Elutrap.

We have made a significant improvement in the electroelution device, Elutrap (Schleicher and Schuell) by substituting an agarose gel barrier, which is made from 0.6% agarose (SeaKem GTG; FMC Corporation), into the elution chamber in place of the manufacturer specified BT2 membrane. This modification substantially increases the DNA recovery from agarose gels, even in samples containing less than 1 microgram of DNA, and shortens elution times particularly for large sizes of DNA (greater than 4.4 kbp). Additionally, the gel barrier provides a reproducible quantity and quality of DNA recovery. The high quality of the eluted DNA using the modified Elutrap makes this system suitable for further DNA manipulations.     

Electroelution of DNA and protein from polyacrylamide and agarose gels

An electroelution method is described for the recovery of DNA and protein from agarose or polyacrylamide gels. The samples to be electroeluted are compartmentalized in a modified microcentrifuge tube fitted with dialysis membranes. This procedure is simple, rapid, inexpensive and efficient. Within 30 min to 2 hrs, the recovery of the sample is nearly quantitative. DNA fragments recovered can be directly subjected to DNA sequence analysis or enzymatic reactions after ethanol precipitation. Proteins can also be recovered after separation by acrylamide gel in the presence or absence of detergents and be ready for further analysis.     

Development and optimization of a binding assay for histone deacetylase 4 using surface plasmon resonance

Electroelution is a widely used methodology for protein purification. In this study, a practical and low-cost system for protein electroelution from stained polyacrylamide gels was developed. For this, a horizontal protein electroelution cuve was constructed with glass plates, 1.5-ml capacity microcentrifuge tubes, and dialysis membrane. Analyses of the system efficiency showed high protein recovery from nonfixed and fixed sodium dodecyl sulfate (SDS)-polyacrylamide gels.     

Purification of plasmid DNA by an improved electrophoretic protocol

Plasmid DNA from Escherichia coli was isolated by electroelution carried out in an agarose gel that contains an incorporated dialysis membrane. As the relative mobility of circular plasmid DNA to linear chromosomal DNA increases when the agarose concentration is decreased, we were able to purify plasmids of up to 50 kbp in 0.3% agarose gel in Tris acetate buffer yielding 10-60 g DNA ml bacterial culture.     

Rapid alkaline transfer of low molecular weight DNA from NuSieve GTG agarose gels

The rapid alkaline transfer of high molecular weight DNA from agarose gels to nylon membranes has greatly decreased the time required for setup of Southern transfers. This technique has been used to resolve genomic DNA greater than 1000 base pairs by conventional electrophoresis on 1% agarose gels followed by alkaline transfer to nylon membrane. Now we report that this rapid alkaline method can be used for the transfer of low molecular weight DNA fragments (10 to 1000 base pairs) from NuSieve GTG agarose gels to nylon membrane.     

Rapid small-scale DNA isolation from filamentous cyanobacteria

Abstract A rapid small-scale DNA isolation procedure is described for the filamentous cyanobacteria, which yields enough chromosomal and plasmid DNA for restriction endonuclease digestions, Souther hybridizations, and electroelution from gels for further manipulation. DNA from seven strains of cyanobacteria were isolated and analyzed on agarose gels.