摘要:This protocol is for Mini (up to 20 µg) preparations of high-copy plasmid DNA from cultures of E. coli .Important notes before startingNew users are strongly recommended to read Appendix A (page[阅读全文:]
摘要:一、原理球状蛋白质一般都可以在低盐溶液或蒸馏水表面形成不溶解的变性薄膜，若浓度合适，则肽链伸展形成蛋白质单分子层。一般用碱性蛋白质包围带负电的核酸分子，当蛋白质展开时，核酸也随之展开，核酸的形态结构将保持一[阅读全文:]
摘要:Procedure1. Grind 2 to 5 g of frozen leaves to a very fine powder with a liquid nitrogen-cooled mortar and pestle.2. Add 25 ml of CTAB Buffer and transfer to a 50 ml tube.3. Incubate at 65℃ for 20 min[阅读全文:]
摘要:1 .核蛋白的提取①用 细 胞 刮子刮取RAW264.7 细胞与VSMC ，移入2.5mlEP 管中。②将细胞用冰冷PBS 洗两遍，于4 ℃ , 5000g 离心3min ，沉淀细胞。③加5 倍细胞体积的冰浴缓冲液A 洗细胞一次，于4 ℃ ,5000g 离心3 min 沉淀细胞.[阅读全文:]
摘要:实验步骤：1．取 10μl DNA 于 0.8% 凝胶检测；2．将 DNA 调节浓度至 300-400 ng/μl；3．仔细阅读将所用的任何一种酶产品说明书，熟悉反应条件及酶切的贮存浓度（ 10U-50U/μl ）厂家配套试剂；4．计算据反应条件所需要[阅读全文:]
摘要:Analysis of Genomic DNA by Southern HybridizationSelect several independent BAC clones containing the same inserts that will be used as a probe. Confirm the BAC clone integrity using by comparing Hind[阅读全文:]
摘要:Digest DNA in 96-well plate To each well add:4μl 10Xbuffer4μl Enzyme0.4μl Spermidine(0.4M)31.6μl H2 O37℃ 19h, then add 4μl loading dye to each well. Load into 400ml 1% agarose gel immed[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:]