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:The Minion Lab, College of Veterinary Medicine at Iowa State University http://mycoplasmas.vm.iastate.edu/lab_site/methods/DNA/SmaIvector.html [阅读全文]

摘要:碱法提取的质粒DNA即使用RNA酶处理，仍会含有少量RNA。当有些试验需无RNA污染的DNA制品时，则需进行进一步纯化。一般常用Sepharose 2B或Sepharose 4B进行纯化，该方法具有快速，条件温和，重复性好，载体物质可以再利用等优[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:1.Take the 384 well viper plate from the thermocycler. Remove the Silicone Sealing Mat (Axygen Scientific: AM-384-PCR-RD) and place face down on a paper towel and pat to absorb any liquid that may be [阅读全文]

摘要:Submission Details: *Dr. Simon Dawson*Department of Biochemistry*University of Nottingham*U.K.*17:03*22/1/96. Database: Method/Protocol-->Spin column purification can be used to change buffers without[阅读全文]

摘要:SNGE,NIMH,NIH http://intramural.nimh.nih.gov/lcmr/snge/Protocols/Miniprep.html SOLUTIONS:•Resuspension buffer: 25mM Tris-HCl (ph8)50mM Na2 EDTA1% glucoseAutoclave and store at 4℃.•NaOH/SDS: [阅读全文]

摘要:Amberg Lab ,Upstate Medical University http://www.upstate.edu/biochem/amberg/protocols/seq_temp.html Notes on DNA : The cleaner the better.If mini DNA use phenol extracted and use the entire mini prep[阅读全文]

摘要:Cepko/Tabin Lab, Harvard University This protocol works well from a 5 ml starting culture or a 1 ml starting culture (adjust volume accordingly).http://axon.med.harvard.edu/~cepko/protocol/mike/S2.htm[阅读全文]

摘要:The standard solution typically used for both pre-hybridisation and hybridisation is based on that given in Maniatis et al.,(1982)with both Denhart's solution and heterologous DNA being replaced by he[阅读全文]

摘要:Cepko/Tabin Lab ,Harvard University http://axon.med.harvard.edu/~cepko/protocol/mike/S6.htmlSuperfrost plus slides and standard slides can be silinized but I have found that superfrost slides work bes[阅读全文]

摘要:1.Pick plaques and put in 500 ml sterile SM with 20 ml chloroform.Store in 4° overnight.2.Be sure we have SOLR and XL1-Blue cells in 10 mM MgSO4,no more than 1 week old.If not,set up a culture of [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]