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摘要: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 [阅读全文]
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摘要:Restriction Enzyme Buffer Most enzymes can use REact buffers; however, some are made up separately. Use fresh Milli-Q water , siliconized or sterile glassware or disposable plastic ware to make the fo[阅读全文]
摘要:Materials:10X restriction enzyme buffer (see manufacturer's recommendation)DNAsterile waterrestriction enzymephenol:chloroform (1:1) 1.Add the following to a microfuge tube:2 μl of appropriate 10X [阅读全文]
摘要:Restriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restriction enzyme, in its respective buffer as recommended by the supplier, and a[阅读全文]
摘要: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 [阅读全文]
摘要:Restriction enzymes are expensive ($40 to $200 a vial): keep the enzymes at -20℃. Plan your digests to be small and convenient. The digest is composed of DNA sample (volume should not be more than abo[阅读全文]
摘要:Probes should be stored frozen both when lyopholized or in solution. Aliquoting your probe into additional vials after resuspension can help to minimize potential breakdown due to repeated freezing an[阅读全文]
摘要: Pat Brown's Lab ,Department of Biochemistry and the Howard Hughes Medical Institute http://cmgm.stanford.edu/pbrown/protocols/amino-allyl.htmA. RT Reaction1. To anneal primer, mix 1-2 m g mRNA with 5[阅读全文]
摘要:The Preuss Lab,The Division of Biological Sciences,The University of Chicago.http://preuss.bsd.uchicago.edu/protocols/enzyme.html[阅读全文]
摘要:H. Simmler and H. SingpielAcconovis GmbHLindenhofstr. 42-4468163 Mannheim, GermanyeMail: simmler@acconovis.comR. M¨annerUniversit¨at MannheimB6, 23-2968131 Mannheim, Germanymaenner@ti.uni-mann[阅读全文]
摘要:Recovering DNA from agarose gelsPaul N. Hengen, Ph.D. (July 14, 1999)IntroductionMethods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.meth[阅读全文]
摘要:I recovered DNA from gel by Millipore's kit,It's simple.Cut the band from gel and centrifuge for 10 minutes,but the efficiency is disappointed,only 20% or so. Can anyone reccomend me other methods?Tha[阅读全文]
摘要:(adapted from Bruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma 73019 broe@ou.edu)Restriction enzyme digestions are performed by incubating double-st[阅读全文]
摘要:The Minion Lab, College of Veterinary Medicine at Iowa State Universityhttp://mycoplasmas.vm.iastate.edu/lab_site/methods/DNA /restrdig.html [阅读全文]
摘要:Recovery of DNA from LMP (Low Melting Point) Agarose Gel1. Separate DNA fragments through an LMP agarose gel containing ethidium bromide (0.5 microgram/ml). 2. Detect DNA by irradiating the gel with l[阅读全文]
摘要:实验中的一些好习惯加入试剂之前,把它混匀一下,以免放置时间长了浓度不均移液枪用完之后要归到最大计量的位置,防止久而久之弹簧失去弹性一定要记着关水浴箱,切记切记多和大家讨论,同时多关注别人讨论的经验,这几乎是[阅读全文]
摘要:1、反应体系 25ulCocktail 20 ul: Primer FP 1 ul + Primer RP 1u + 2 * SYBR GREEN I MIX 12.5 + H2O 5.5 ul cDNA 5 ul2、Primer 浓度,需要摸索,从200nM到700nm等,设置不同浓度。3、每个引物浓度,从well 1到12设置4个样[阅读全文]
摘要:Q: Does the PCR reaction volume (negatively) influence the outcome?A: No, especially since the introduction of the small, thin walled, 0.2 ml plastic vials fitting the 96 well metal blocks of the ther[阅读全文]