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: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: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: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: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]

摘要:DNA PrepPrepare DNA via your favorite method.You may find a protocol underMini Yeast Genomic Prep.Restriction Digest1.Digest DNA with appropriate restriction enzyme.Digest 10μg of Yeastchromosomal [阅读全文]

摘要:1. Run the gel as normal. Often for genomic southerns it is desirable to run long gels (18cm) over 4-6hrs.2. Photograph the gel with a ruler adjacent to the molecular weight markers as a reference.3. [阅读全文]

摘要:核酸分子杂交技术用一段核酸片段带上特殊标记物后，在一定温度，离子强度，PH条件下，与相同or相似的核酸分子以硷基互补原则（C≡G, A＝T,A＝U）形成新的杂合分子的过程，以检测核酸分子的存在。根据反应性质分为：RNA-RNA, RNA-[阅读全文]

摘要:PCR screens must be designed to detect transgene DNA at the single copy level. Southern Blots analysis of transgenic mice need copy standards to estimate copy number. Copy standards are prepared by mi[阅读全文]

摘要: DNA的提取与检测人和哺乳动物细胞基因组DNA的分离通常是在有EDTA、Sarkosye等一类去污剂存在下，用蛋白酶K消化细胞，随后用酚、氯仿抽提，经RNase处理和纯化来提[阅读全文]