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原理 DNA 以核蛋白形式存在于细胞核中，制备 DNA 的原则是既要将 DNA 与蛋白质、脂类和糖类等分离，又要保持 DNA 分子的完整。蛋白酶K 及链霉蛋白酶 E 的应用使这两个原则得到了保证。 蛋白酶K 或链霉蛋白酶 E 均为广谱蛋白酶，其重要特性是能在 SDS 和 EDTA 存在下保持很高的活性。在匀浆后提取 DNA 的反应体系中， SDS 可破坏细咆膜、核膜，并使组织蛋白与 DNA 分子分 ...

Use from 0.01 - 0.1 gram plant material. Grind the plant material with liq. N2 in a mortar. We normally use some alumina to crush hard tissue. Transfer the ground tissue to a eppendorf tube. Add 1 ...

This clearly is an "old procedure revisited" but is extremely efficient because 1) the amount of resulting nested fragment set is at least 5 to 10 times more than obtained by Sephadex G-50 ...

Yale Genome Analysis Center Yale University http://ygac.med.yale.edu/mtn/LS_transf_prot.stm The following protocols were developed by Petra Ross-Macdonald for Mike Snyder's LacZ fusion project at Yale ...

Procedure 1. 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 a ...

I am seeking procedures and/or advice on how to isolate human genomic DNA from platoregistry sera. The sera is lymphocytes that have been frozen for 35-40 years but are not dessicated. Will the isolat ...

1. Run gel ...

Dige ...

Pat Brown' ...

1) Digest DNA with two restriction enzymes such that one end is ExoIII susceptible (blunt or 5' overhang) and the other is ExoIII resistant (5' recessed). The ExoIII susceptible end should be such tha ...

Analysis of Genomic DNA by Southern Hybridization Select several independent BAC clones containing the same inserts that will be used as a probe. Confirm the BAC clone integrity using by comparing Hi ...

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的各种试剂准确用量：（ 0.5 ml tube 中） DNA(3-5μg)&nb ...

http://axon.med.harvard.edu/~cepko/protocol/mike/F1.html This protocol was designed to generate directionally end-labeled probes for DNaseI footprinting but it can be used for any application that re ...

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 HYBRID ...

This protocol is for Mini (up to 20 µg) preparations of high-copy plasmid DNA from cultures of E. coli . Important notes before starting New users are strongly recommended to read Appendix A (pa ...

Dear ...

DNA的重组 DNA Recombination一、同源重组 发生在同源序列间的重组称为同源重组(homologous recombination)，又称基本重组。 是最基本的DNA重组方式，通过链的断裂和再连接，在两个DNA分子同源序列间进行单链或双链片段的交换。 二、细菌的基因转移与重组（一）接合作用(conjugation)： 当细胞或细菌通 ...

Isolation of DNA from Mouse Tail Biopsies 1. Obtain tail biopsies from 2 to 3 week old mice: Hold mouse firmly at base of tail with one hand with the other cut off 0.5 to 1.0 cm of the tail tip with ...

Preparation of Frozen Competent E. coli 1. For TG1 or DH5a cells start with a colony from a minimal plate to ensure the presence of the F ' episome. Grow in 5 mls LB o/n (see note on medium below). In ...

Hidoes any ...