DNA提取与纯化

1. Remove 0.5 cm of tail into polypropylene microfuge tube (do not mince). (The tubes must have tight-fitting caps so that there are no leaks in steps 3 and 7 below.) 2. Add 0.5 ml DNA digestion buffe ...

This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure with a variety of Populus species, as well as tobacco and Arabidopsis . The resulting DNA is of sufficiently high quality for PCR (including RAPD), restriction digests, and ligation reactions. However, it is extemely important to use the youngest leaves available for Populus , as DNA from older leaves is often contaminated with compounds that can interfer

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Materials: Silica Suspension: add 2 g of silica to 15 ml of H2O wash 3x by centifugation at 2000 x g for 2 min estimate vol of silica and resuspend in 2 vol H2O Silica Wash Solution: 50 mM NaCl, 10 mM Tris 7.5, 2.5 mM EDTA, 50% Ethanol 6 M NaI

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The following protocol is based on our modifications of R. Kraft, J. Tardiff, K. S. Krauter, and L. A. Leinwand. Biotechn . 6(6):544-545, 1988. Inoculate 2-5 ml of L broth containing the appropriate antibiotic from a single bacterial colony. Incubate at 37℃ overnight with vigorous shaking. Follow the Plasmid Quick Prep protocol through the potassium acetate step to isolate plasmid DNA. Add DNAse-free RNAse to 50µg/ml, incubate, 37℃ from 10-30 minutes. Phenol extract the solution, saving the uppe

一、DNA 提取　 SDS 高盐法(方案1) 具体步骤: 称取1 g 土壤，放入研钵中，倒入适量的液氮，立即研磨；再倒入适量的液氮，研磨，重复3 次，使土壤颗粒研成粉末； 将13.5 ml 提取缓冲液(0.1 mol/L磷酸盐 ，0.1 mol/L EDTA ，0.1 mol/L Tris-HCl ，1.5 mol/L NaCl ，1.0 % CTAB) 和50&mu ...

Materials: 1.TENS solution: 10 mM Tris (pH to 7.5) 1 mM ethylenediaminetetraacetic acid (pH to 8.0 to dissolve) 0.1 N sodium hydroxide 0.5 % sodium dodecyl sulfate 2.3 M Sodium acetate pH 5.2 3. ...

1.grow plants in trays of 96 and leave two spots open (for the PCR controls) 2.harvest 1 to 2 young and green leaves (1cm2 /plant at rosette stage if possible). Use 96 well plates (1 or 2 ml E&K ...

CTAB TECHNIQUE / Method / Schedule / Protocol (JPB) FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES (see also DNA RNA double isolation procedure if both DNA and RNA are needed) Rea ...

Preparation of BAC (Bacterial Artificial Chromosome) DNA with CONCERT ™ High Purity Plasmid Purification System1 Courtesy Lisha Xu and Alice C. Young Molecular and Cell Biology Research and Deve ...

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Important : Extract the DNA within one week of receiving samples. Samples that have been processed should be frozen to prevent degradation. Freeze samples between use each day. 1. Add 600 m l of 50 mM NaOH to a 1.5 ml eppendorf tube and insert brush in tube. (Clip off handle of brush with wire cutters so tube can be closed. Cutters should be rinsed with EtOH between samples to prevent contamination.) 2. Vortex thoroughly to mix, for at least 10 s. 3. Heat to 95℃ for 5 min. 4. Spin briefly to poo

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1取5ml噬菌体溶液放入无菌干净的小烧杯中，缓缓加入固体NaCl，使其终浓度为1M（0.292g）充分溶解后冰浴1小时。 24℃下11000rpm离心10min，取上清液于另一个干净烧杯中，将PEG6000加入烧杯中使其终浓度为10%W/V约0.5克），磁力棒缓慢搅拌，冰水浴1.5小时，于4℃，15000rpm离心15min，弃上清。 3将沉淀溶于300μlTE（pH8.0），加等体积的酚/ ...

DNA 是遗传信息的载体，是最重要的生物信息分子，是分子生物学研究的主要对象，因此DNA 的提取也应是分子生物学实验技术中的最重要、最基本的操作，如不能有效的完成DNA 提取方面的工作，那就根本谈不上进行分子生物学方面的实验。在DNA 提取过程中应做到1，根据不同研究需要，保证结构的相应完整性；2，尽量排除其它大分子成分的污染(蛋白质、多糖及RNA 等)；3，保证提取样品中不含对酶有抑制作用的有机 ...

1.Obtain 65-100 µl of blood by retro-orbital bleed with a heparinized microcapillary tube. Expel blood immediately into a 1.5 ml microfuge tube containing 20 µl of 10 mM EDTA. Mix immediat ...

Materials: phenol:chloroform (1:1) chloroform Add an equal volume of buffer-saturated phenol:chloroform (1:1) to the DNA solution. Mix well. Most DNA solutions can be vortexed for 10 sec except for high molecular weight DNA which should be gently rocked. (If using Phase-Lock Gel, follow procedure M.1 ) Spin in a microfuge for 3 min. Carefully remove the aqueous layer to a new tube, being careful to avoid the interface. (Steps 1-4 can be repeated until an interface is no longer visible) To

Materials: • 0.8 % agarose gel in 1x TAE • Digested DNA • Glass Milk • NaI solution • New Wash Procedure: 1) Run digested DNA out on agarose gel slowly (70 V on BioRad gel) 2) Use long wave UV lamp to visualize bands. Cut out band with scalpel. Cut smallest possible piece. 3) Put gel slice in an eppendorf tube and weigh to figure out volume of gel slice. (empty tube approx. 1 g). 4) Add 3 vol of NaI solution (gel slice is usually ~200 mg, therefore, add 600 µl NaI). 5) Melt gel slice in 55℃ wate

常规片断级的选择指南： 所谓常规级，即回收片段大小介于100bp和10kb之间，在这个范畴内包含了一般的质粒或者克隆片段的回收。主流方法是柱回收试剂盒。 1. Qiaquick Gel Extraction Kit 品牌：Qiagen 回收范围：70bp-10kb 特点： 高达80%回收率 操作快速简单，3步完成70bp-10kb片段的回收 提供三色指示剂的电泳上样loading Bu ...