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1.显性标记与选择效率 一部分分子标记系统如RAPD、AFLP具有显性或部分显性的特点，无法区分基因是纯合还是杂合，不能提供完整的遗传信息。Haley等（1994）提出相斥相（Repulsion－phase）的RAPD标记无论对显性还是隐性均可极大地提高选择效率，他们找到了和菜豆普通花叶病毒抗性基因bc-3连锁相引标记-1，相斥标记bc-2，用标记-1选择纯合抗病株、杂合体、纯合感病株分别占26. ...

磷酸钙-DNA 共沉淀法 核酸以磷酸钙-DNA 共沉淀物的形式出现时，可使DNA 附在细胞表面，利于细胞吞入摄取，或通过细胞膜脂相收缩时裂开的空隙进入细胞内，进入细胞的DNA 仅有1%～5%可以进入细胞核中，其中仅有不到1%的DNA 可以与细胞DNA 整合，在细胞中进行稳定表达，基因转导的频率大约为10-4，这项技术能用于任何DNA 导入哺乳类动物进行暂时性表达或长期转化的研究。此方法对于贴壁细 ...

1.如何测定引物的OD值？ 用紫外分光光度计在260nm波长测定溶液的吸光度来定量，请注意紫外分光光度计的使用，测定时溶液的吸光度最好稀释到0.2-0.8之间(吸光度太高或太低会有较大的误差)。DNA 干粉用一定体积的水充分振荡溶解以后，取部分溶液稀释到1ml并在1ml标准比色皿中测定其吸光度，即为所测体积的OD值，进而可以计算出母液的OD值。 举例：您拿到一管干粉的DNA ，用1ml水溶解成 ...

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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I am having huge problems with bisulfite PCR. Southern analysis indicates that my DNA is unmethylated. My first bisulfite treatment also indicated that the DNA was unmethylated. Subsequent treatments ...

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David Harry Institute of Forest Genetics USDA Forest Service Pacific Southwest Research Station August 26 1993 Background : There are many published methods for mini-preps of DNA from lambda phage cl ...

Inoculate a 5ml overnight of E.coli in LB+20 mM MgSO4. Next morninginoculate 250 ml LB+20 mM Mg++ in a 2L flask with about 2ml overnight culture.Grow at room temp (23℃)with good aeration (250rpm)to an ...

SOLUTIONS: Green and blue solutions from Clone Checker kit (cat. no. 11666-013). PROCEDURE: Place 3-6µl of an overnight culture in an Eppendorf tube (or spread part of a bacterial colony on the ...

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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 Yeastchromosoma ...

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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- ----------------------------------- ...

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 ...

Hey I am analyzing the methylation status of two regions: one ecompasses 500bp the other is around 800bp. Could I just designed primers at flanking sites and amply these two regions? I found usually o ...

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Jacks Lab Center for cancer researchMIT 1. Each tail should be in a clean eppendorf tube. 2. Add 500µl of tail lysis buffer containing Proteinase K (PK) to each tube. 3. Incubate tai ...