DNA提取与纯化

This protocol gives very clean plasmid preps for restriction digests and cloning. However due to the alkaline lysis step the DNA is often nicked and may not give exceptional sequence data.SolutionsTEN ...

Materials:3 M sodium acetate pH 5.2 or 5 M ammonium acetate DNA 100% ethanolMeasure the volume of the DNA sample. Adjust the salt concentration by adding 1/10 volume of sodium acetate pH 5.2 (final ...

简介 优点：操作简单方便 缺陷：不能像EMSA或footprinting一样区分不同类型复合物 原理 双链 DNA 能够通过硝化纤维膜 (NC)，但是 DNA-protein 复合物不能通过NC filter. 通过定量分析留在滤膜上的复合物，可以判断二者的结合常数。 基本方法Lable DNA Mix DNA and protein to form complex Pass the mixtur ...

Pyrosequencing 在肿瘤基因甲基化研究的应用 甲基化研究对于基因的调控和肿瘤的发生有非常密切的关系。在一般正常的细胞中，基因调控区CpG岛处于非甲基化的状态。而当细胞发生癌变后，这些CG区域往往呈现甲基化状态。英国医学刊物《Lancet》报道奥地利因斯布鲁克医学院的专家们早就可以通过检测大便中DNA的甲基化变化，把健康人的大便与结肠癌患者的大便区分开。大便标本中SFRP2 甲基化的程度 ...

Molecularpourous membrane tubing is used for desalting protein and DNA isolation / purification. It comes in several diameters and pore sizes (for molecular weight conversions for nucleic acids see ge ...

Purification of DNA from agarose gels is an essential method involved in the sub-cloning of DNA fragments. The following method describes a variation of the method of Vogelstein and Gillespie 1979 (Pr ...

Pellet 1.5 ml of an overnight culture at 12000 rpm in Eppendorf centrifuge at RT for 3 min. Resuspend bacterial pellet in 350 µl of STET buffer. Add 25 µl of freshly prepared solution of lysozyme (10 ...

1.Obtain 65-100 µl of blood by retro-orbital bleed with a heparinized microcapillary tube. Expel blood immediately into a 1.5 ml microfuge tube containing 20 µl of 10 mM EDTA. Mix immediately to preve ...

This protocol is for Mini (up to 20 µg) preparations of high-copy plasmid DNA from cultures of E. coli.Important notes before startingNew users are strongly recommended to read Appendix A (page 21) at ...

1、将已经电泳确定的可回收的酶切产物在合适浓度的回收用琼脂糖凝胶进行电泳。最好换用新的电泳缓冲液10 × TAE（10 ×Tris-乙酸）。2、当溴酚蓝迁移至足够距离时（至少2 cm以上），在长波紫外灯下观察，用清洗过的刀片在目的片段前切下与目的片段同长，宽度适当（一般2 cm左右）的胶块。注意：①不要忘记在胶下垫一个新的塑料手套防止污染。②小心不要将回收胶切裂，同时注意与目的带相邻的切面要尽量平 ...

1. Prepare Whatman DE-52 according to manufacturers specifications. Equilibrate at store with 0.02% Sodium azide.2. Plug a 1 mL (blue) pipet tip with Siliconized glass wool. Add 500 ml of 50:50 DE-52 ...

PurposeThe protocol describes how to prepare human tonsil lysate for use in purification of ICAM-1LFA-1and PNAd.MaterialsSafety EquipmentsLab CoatLatex GlovesFace MaskBench PaperFresh human tonsil tis ...

Does anybody know how to isolate single-stranded DNA from mammalian cells?Thanks for responding.Yin----------------------------------------------------------------------------------HelloI don't know i ...

Hattoti's protocol adapted to cell culture :Hattori MTugores AVeloz LKarin MBrenner D (1990)A simplified method for the preparation of transcrip-tionally active liver nuclear extracts.DNA Cell Biol.Vo ...

NH4Ac and EtOH precipitation of DNA Add NH4Ac (10M stock or solid) to the sample for a final concentration of 2.5M mix (spin at 4℃ transfer the supernatant to a new tube; optional spin for extra purif ...

PROTOCOL TO EXTRACT DNA FROM PARAFFIN BLOCKS1.Cut 10-20X10μm sections of formalin fixed paraffin samples into eppendorf tubes.2.Add 1 ml xylene mix incubate at 55℃ for 15mins. Release pressure spin ...

CTAB TECHNIQUE / Method / Schedule / Protocol FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES(see also DNA RNA double isolation procedure if both DNA and RNA are needed)Reagents ne ...

DNA Fragment Purification from Agarose or AcrylamideFor fragments from 200 bp to 10 kb the agarose purification is ideal.For smaller fragments (20 bp to 400 bp)the acrylamide purification is preferred ...

Source: Contributed by Mohammad Reza Abbaszadegan et al. Abstract: Single strand conformational polymorphism (SSCP)is the most widely used PCR-based methods for point mutation detection. The abnormal ...

1、Oligo design（1）The 5' end (the homology arm) - choose 42 or more (we usually choose about 50) nts for the homology arms from the target DNA sequence simply according to where you want to insert the ...