DNA实验技术

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 ...

人类基因 ...

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 ...

方法一 A液：1M，MnCl2： B液：1M，HEPES，pH=6.2-6.8，用无菌水配，配后不需灭菌； C液 称取CaCl2 0.10g，KCl 1.18g，全部转入细口试剂瓶，然后加入46ml三蒸水，轻轻振荡使所有组分充分溶解。将瓶塞盖上并用牛皮纸、棉线包扎，然后放入灭菌锅121℃高压灭菌备用。 取1管B液（0.6ml），全部加入C液(46ml)中，混匀后，再用1ml移液器加入3.3m ...

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. ④加3 倍细胞体积的冰浴缓冲液A 悬浮细胞，冰上放置30min 、剧烈振荡10 次，4 ℃ 5000g 离心15min ...

准备： 1．灭菌 试管 400毫升细长烧杯2瓶，离心瓶4-6个（250ml）。 2．试剂：YEP 1200ml（每瓶300ml 共4瓶） Kan 1;1000，Rif1:500。 1/2MS 2%蔗糖（灭菌115度20分钟），Silwet在-20℃贮存。 3．步骤： 共转化农杆菌：于中午12点接菌于有YEP培养液的试管中10ul：10ml接种。28℃，3000rpm摇过夜，约30小时，次日下午6点 ...

1. Run gel ...

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 ...

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 ...

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 ...

Pat Brown' ...

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 ...

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 ...

Dige ...

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 ...

1．如何做酶切反应？ 该问题看似什么简单： DNA 中加上酶，然后保温一段时间就可以了。但是在实际操作过程中，我们不断听到：切不动，装不上。问题在什么地方？能系列生产限制性内切酶的公司国际上，就那么几个，位列前3 的是NEB Fermentas SibEnzyme。这些公司提供酶的品质一般都能得到保证。您可以怀疑酶的质量问题，但是更多的问题来源于模板是否合适酶切要求。下面几点对你的酶切是有帮助的 ...

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 ...