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搜索“TAG:Southern”找到相关内容27篇,用时0.018341秒
摘要:基本原理是:硝酸纤维膜或尼龙滤膜对单链DNA的吸附能力很强,当电泳后凝胶经过DNA变性处理,覆以上述滤膜,再于其上方压上多层干燥的吸水纸,借助它对深盐溶液的上吸作用,凝胶上的单链DNA将转移到滤膜上。[阅读全文]
摘要:1. Run gel (0.8 -1.0% agarose is best). For yeast chromosomal Southerns, digest 20 μg DNA. Use 50 ml minigel for most purposes. Photograph gel, but minimize exposure to UV light.2. Depurination: Pl[阅读全文]
摘要:Southern Hybridization Protocols Two protocols are given here.The first one is used for BAC library screening on filters, although is also suitable for Transfer hybridizations. PREPARATION OF HYBRIDIZ[阅读全文]
摘要:Hi can anyone answer thisI am having problems with my southern hybridization.I am wondering if the DNA transfers to my membrane.Is it possible that the DNA can wash into the solutions when denaturing [阅读全文]
摘要:一 酶切和电泳在200 μl 微量离心管中加入:25 μl DNA样品(约10μg),3μl 限制性内切酶(MBI,10 U/ μl)5 μl 相应的10×buffer,补水到50μl。然后加一滴矿物油覆盖, 稍微离心后放于37℃水浴8-12小[阅读全文]
摘要:This kit contains materials for six groups to perform Southern transfer and hybridization analysis using the included lambda DNA samples and biotinylated probe. The intellectual objective of the exper[阅读全文]
摘要:Has anyone any experience of using a semi dry blotter for Southern blotting. At present I am tranfering overnight using a weight, I think there is a blotter that is suitable for nucleic acids and can [阅读全文]
摘要:1) 取10μl待测DNA,于一定浓度的琼脂糖凝胶上进行电泳。(20mL,1.0%凝胶)2) 凝胶用溴化乙锭染色,切掉凝胶四周多余部分,并在凝胶的一角作一记号, 拍照记录电泳结果(拍照时, 凝胶旁放一尺子)。3) 杂交用胶的制备 (做以[阅读全文]
摘要:Two protocols are given here. The first one is used for BAC library screening on filters , although is also suitable for Transfer hybridizations. PREPARATION OF HYBRIDIZATION SOLUTION Prehybridization[阅读全文]
摘要:Materials:Whatman 3 mm Blotting Papernitrocellulose (Schleicher & Schuell, Amersham) or nylon membrane filter (Amersham).Paper towels (preferably C-fold "cheap-o" variety)Pyrex or Tupperware dish, gla[阅读全文]
摘要:Day 1 1. Digest DNA for 6 hours (or overnight)BSA 10 mg/ml 0.5 22.65 μl of 10 μg Genomic DNARnaseA 10mg/ml 0.1 in TE mixed to 7.35 μl cocktailSpermidine 100nM 0.75 per sample10X enzyme buffer[阅读全文]
摘要:Southern Blotting: Detection via Chemiluminescence (BM's Genius System) Modified to include the use of CSPD for chemiluminescent detection! This is the method we are currently using for Southern blots[阅读全文]
摘要:1. Depurination of DNA fragments: Wash gel in 0.25 M HCl 5' (small gels), 7' (large gels)2. Denaturation: Wash gel in (0.5 M NaOH, 1.5 M NaCl) for 20' (small) to 30' (large).3. Neutralization: Wash in[阅读全文]
摘要:-Elaine Pinheiro and Aurora Burds ConnorMIT, Jan2006This protocol works very well with Hybond-XL membrane from GE Healthcare (formerly Amersham).1) Digest your genomic DNA, using about 10 μg per re[阅读全文]
摘要:I'm trying to do a southern blot and I have a trouble, the bands I get are the same bands that appear in the genomic DNA digestion, my probe is specific and it works in northern blot.Thanks-PPlopez---[阅读全文]
摘要:Hi everyone,i'm looking for a good southern blot protocol with all the secrets of this technique. I never did it but I know is not too difficult. I'd like a protocol that show me the "way to do" fro d[阅读全文]
摘要:This is a brief overview of how a Southern blot (more formally called an DNA blot)is performed and what type of data you can obtain form one.Figure 1.Southern blots allow investigators to determine th[阅读全文]
摘要:DNA Transfer1. Depurination of DNA fragments: Wash gel in 0.25 M HCl 5' (small gels), 7' (large gels)2. Denaturation: Wash gel in (0.5 M NaOH, 1.5 M NaCl) for 20' (small) to 30' (large).3. Neutralizat[阅读全文]
摘要:1.Digest DNA: • 1.5 μg DNA in a 25 μI reaction containing BSA• 3 to 5 units for 5 to 3 hours (aim for 10-fold overdigestion; remember many enzymes will not survive well for more than 3[阅读全文]