Southern Blotting

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 ...

Transgenic Mouse and Gene Targeting Facility Tg 008 Southern Blotting Materials and Equipment Hybaid hybridization oven (ISC BioExpressH-9250) Vacuum oven (VWR52201-218) Ludlum Geiger Counter Radiati ...

Hi Can you tell me what do "SSC" in a southern blot how he act on DNA or in the protocole. (excuse-me for my traduction i never write in english) -Sebela- ----------------------------------- ...

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 ...

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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 ...

I am trying to digest mouse genomic DNA for a southern blot. When digested with either HIndIII or NdeI I get a nice smear as expected. But when I digest with either speI or BglII (neither one of which ...

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 hep ...

对于大的基因组，DNA 酶切图谱凭肉眼是分辨不开的（EB 染色），因为大小不等的分子呈现弥散分布，只有借助灵敏的放射性同位素（或其他化学发光物质），将靶 DNA 在凝胶上（膜上）的带型通过特定的探针与之杂交，转换成 X 光片上直观的带型，才能进行相关分析。另外如果需要鉴定或寻找与已知 DNA 同源的 DNA 片段如：染色体步查、基因组文库的评价和利用、阳性克隆的分析鉴定、转基因拷贝数分析等也都需进 ...

RNA/Zeta Probe Dot Blotting protocol 1)Make up RNA (up to 20μg)dissolved in sterile H2 OTE or 0.5% SDS to 500μl with ice-cold sterile 10mM NaOH1mM EDTA and apply it to Zeta Probe membraneheld in ...

Solutions Protocol: Digest 5-10 μg genomic DNA overnight with restriction enzyme of choice. Run digested gDNA on 0.8% TAE gel with marker (with no ethidium bromide). Transfer Setup: 1. Remove gel ...

Non-radioactive Probes I. Via random hexamers 1. Solutions: 10X hexa nt miundefined: 500 mM Tris-Cl pH 7.2 100 mM MgCl2 1 mM dithioerythritol (DTE) 2 mg/ml BSA 62.5 A260 units/ml (1.56 mg/ml) random hexan ...

DNA 分子是分子生物学研究的基本材料，依不同的实验目的可采取不同的抽提方法获取数量和质量不等的 DNA 。 实验目的：了解植物 DNA 抽提的主要方法，掌握 CTAB 法快速抽提水稻 DNA 。 实验材料及试剂： 水稻叶片，1.5 × CTAB ，氯仿 / 异戊醇 (24:1) ，95% 乙醇或无水乙醇等 实验原理：植物 DNA 的抽提常采用两种方法： （1）SDS 法： 离子去污剂 ...

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& ...

Author: Suzanne Gerttula Date: March 141994 1.Digest between 1 and 10 μg Genomic DNA. I digest at 37 C for about six hours over the course of a day. Spin down briefly heat to 65℃ for 10 min ...

转膜是把DNA 从琼脂糖凝胶中转移到固相支持物（一般是尼龙膜）上固定，是进行各种后续研究（如 RFLP 分析，阳性克隆的筛选验证等所有涉及分子杂交的研究）的前提。 实验目的： 掌握 Southern blotting 的原理及操作步骤 实验原理： 1．转膜的方式： 向上的毛细管转移 向下的毛细管转移 同时向两张膜转移 电转移 真空转移 ...

About 1/2 to 1 cm of tail should be cut from properly marked mice (using toe or ear clipping etc) about the age of 2-3 weeks. Place these tails into marked 1.5 ml Eppindorf tubes for processing; they ...

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- - ...

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 blo ...

-Elaine Pinheiro and Aurora Burds Connor MIT Jan2006 This protocol works very well with Hybond-XL membrane from GE Healthcare (formerly Amersham). 1) Digest your genomic DNA using about 10 μg per r ...