DNA实验技术

PURPOSE: TolocateaparticularsequenceofDNAwithinacomplexmixture(locateonegenewithinanentiregenome) SeparatemixtureofDNAsequencesonagelprobewithspecificDNAsequence(gene) Otherrelatedblottingtechniques: ...

1. Add an equal volume (equal to sample volume) of P/C to sample.2. Mix (shake don't vortex).3. Take aqueous (upper) layer. (If dirty sample repeat Ph/Chl step until interface is fairly clean).4. Add ...

IntroductionMethods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methds-reagnts available on the internet. A commonly occurring theme on t ...

This is a scaled up version of the Baron protocol which has been modified to achieve purity comparable to CsCl preps. I have not sorted out strain variations yet but HB101 works well.1、SolutionsSoluti ...

There are many methods for DNA preparation without using kits.Below are a few minipreps.These protocols may be scale up to midi or maxi prep if necessary.1.Ammonium Acetate Method 1)Spin down cell and ...

1.Prepare or obtain Buffered phenolpH 8.0.Add 2 crystals of 8-Hydroxyquinoline to prevent oxidation.This also identifies the organic phase as yellow-colored.2.Combine DNA sample with an equal volume o ...

1、Grow an overnight culture from a single colony in 2 mls LB antibiotic.2、Transfer 1.5 mls of culture to eppendorf tube.Spin for 1 minute on high and remove supernatant.3、Add 700 µl STET. Add 25 µl ...

1、Put a 0.5-1.5 CM MOUSE TAIL SNIP in a 1.5 ml eppindorf tube and store frozen until digestion.Make certain that the eppindorf tubes seal wellotherwise your samples may leak out during the shaking ste ...

Sample Preparation Cheek cells are obtained by rinsing the mouth with 25mls of any commercial mouth wash solution available for about 30sec the first thing in the morning. It is important not to brush ...

1.grow up single colony in 1.5 ml LB/antibiotic ovn @37℃ .2.pour into tube spin for 30 sec .3.decant supernatant and resuspend pellet in 100 ml lysis solution .4.add 200 ml alkaline SDS vortex well .5 ...

This is a standard large scale prep.for plasmid DNA which gives a yield of 0.5-1.0mg.I have made some minor changes to the MHB protocol.Solutions Solution IIIIII from protocol D.1.Tris/EDTA pH 7.5 (op ...

A. Culture Growth: 1.For BAC isolation for shotgun library construction: Pick a smear of colonies and inoculate 3 ml TB medium containing the appropriate antibiotic. Grow the culture for 8 hours 37 de ...

1、The BAC clone from glycerol culture sublibrary is innoculated into 3 ml of LB with 20µg/ml chloramphenicol and cultured at 37°C with shaking for 16hrs. The liquid bacterial culture is transferred in ...

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After an aliquot of the PCR mixture is analyzed on an agarose gel the remainder of the reaction is concentrated by ethanol precipitation resuspended in buffer and subjected to a simultaneous fill-in/k ...

1、A smear (rather than a single colony) of BAC colonies are picked and transferred into a 12X75 mm Falcon tube containing 3 ml of LB medium (10 g Bacto-Tryptone 5 g Bacto-yeast extract and 10 g NaCl i ...

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 ...