摘要:Steven Finkbeiner,Departments of Neurology and Physiology,UCSF http://gweb1.ucsf.edu/labs/finkbeiner/Protocol/protocol_list/cloning.shtml DIGEST:1.Digest 10 - 20μg DNA to isolate either the desired[阅读全文]

摘要:1.Wash the plates briefly using the 96-channel block washer. 2.Place the plates in a tub submerged in dd-water. 3.Put a flask on top of the plates to keep them submerged. 4.Place the tub with the subm[阅读全文]

摘要:hi there,I have been trying to clone 3.4 kb insert into pcDNA3.1- for months without any success.The insert is subcloned in TOPO from where I cut it out by KpnI and NotI and purify it by gelelectropho[阅读全文]

摘要:This procedure was originally developed for Listeria monocytogenes but has worked well with other Gram+ bacteria we've tried.1.Pellet cells from 10 ml overnight cultures in BHI or LB and wash in 5 ml [阅读全文]

摘要:Hi,I looked for CpG islands within the promoter of my gene.MethPrimer and CpG plot analysis revealed that in this genes promoter region there were two big CpG islands at locations 280-603 and 693-859.[阅读全文]

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摘要:用含氯化铯和溴化乙锭的悬浮密度梯度离心法分离质粒和染色体DNA的方法，取决于线状DNA和闭环DNA结合的溴化乙锭的量的差异。 [阅读全文]

摘要:通过不连续或预先形成的氯化铯（CsCl）梯度离心，可在离心管中获得含有不同浓度CsCl的溶液分层，样品位于中层（三级梯度法）或底层（两级梯度法）。在随后的离心梯度梯度形成过程中，DNA找到其等密度点，从而使离心时间大大缩短[阅读全文]

摘要:本课件介绍了克隆载体类型、特点及载体必不可少的基本元件。本篇文章来源于网络，如有异议请联系我们，我们将在3个工作日内作出处理。[阅读全文]

摘要:致癌物促进CpG或MpG脱氨基；MpG脱氨基的速度高于CpG：PyMpG发生C-T转换的频率是PyCpG的10倍；非CpG岛区的散在CpG位点为MpG：MpG脱氨基主要发生在非CpG岛区；MpG为突变热点：30％的p53基因点突变发生在MpG位点上 。本篇文章来[阅读全文]

摘要:CONCERT细胞质RNA 纯化试剂是一种新颖的单相酚溶液，是专门设计用来从新鲜或冷冻的培养的动物细胞 或组织中快速、简单地纯化高质量的细胞质RNA 。应用利用CONCERT™细胞 质RNA 试剂分离的总RNA 能够用于构建cDNA文[阅读全文]

摘要:Promega公司的RibocloneR M-MLV(H- ) cDNA合成系统采用M-MLV反转录酶的RNase H缺失突变株取代AMV反转录酶，使合成的cDNA更长。该系统的第一链合成使用M-MLV反转录酶，cDNA第二链合成采用置换合成法，采用RNaseH和DNA聚合[阅读全文]

摘要:First strand of cDNA synthsis1.Add 2 μl of Not I primer-adapter to a sterile 1.5 ml RNAase free tube, add 4.5 μl mRNA of molt4 cells (~5.2 ug), add 0.5 μl DEPC-H2O.2. Heat the mixture at 70℃ [阅读全文]

摘要:CreatorTM SMARTTM cDNA Library Construction Kit整合了SMARTTM技术和与BD Clontech’s pDNR-LIB载体，该载体采用Creator重组技术，提供无需连接接头，直接克隆的快速方法。本篇文章来源于网络，如有异议请联系我们，我们[阅读全文]

摘要:1. Primers:i) Oligonucleotide primers are generally supplied as "so many OD units/ml" - but what does this mean, in terms of mg/ml, or mmol/ml, etc?Given: a primer is Y nucleotides (nt) long;Given: th[阅读全文]

摘要:In designing primers for PCR, the following steps/rules were tested and proven to be useful:1,Length of individual primers between 18-24 bases. Longer primers (30-35 bp) seem to work in more similar c[阅读全文]

摘要:PCR:We generally use 1-2 ul of starting paraffin microdissected DNA for each 50 ul DOP-PCR reaction.We assume that about 1 ug of product is produced in each DOP-PCR reaction. The entire product is the[阅读全文]

摘要:This procedure will work for both yeast and E. coli:Take a small colony and suspend it in 5ul of H2O in a PCR tube. Heat for 5 min at 95℃ and then spin the condensation down in a microfuge. Set up the[阅读全文]

摘要:A method to re-PCR unique bands from products of mixed sizeINTRODUCTIONThe products of a PCR reaction - especially when this is done on eukaryotic genomic DNA, and when using degenerate primers - ofte[阅读全文]

摘要:Reagents / Solutions Lysis Buffer:10ml 10% Sodium dodecyl sulphate (SDS)10ml Glycerol10ml b-mercaptoethanol8ml 0.5M Tris pH6.81ml 0.1% bromophenol blue51ml H2OProtocolSpin down 106 cells and wash, if [阅读全文]