DNA提取与纯化

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定的完整性。然后将核酸分子捞到支持膜上，经过负染色，就可以在电镜下观察。 二、目的 观察质粒DNA 在电镜下的形态，掌握质粒DNA 的电镜制样原理和方法。 三、材料、试剂和仪器 1、纯化的质粒 ...

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 ...

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 ...

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 ...

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 ...

Phenol (removes protein) 1.add equal volume of Phenol (= tris-saturated Phenol-Chloroform-Isoamyethanol) 2.vortex 3.spin 2 minutes at 12000 rpm 4℃ 4.transfer supernatant to a fresh tube (avoid aspi ...

11、将树脂/DNA 混合液抽干后加13ml柱子洗脱溶液至离心管中 对管底部的树脂/DNA 进行洗脱(柱子一边旋转一边加入洗脱液)，并加入柱子中。 12、真空抽干所加入的洗脱。 13、 再加12ml柱子洗脱液进柱子并抽干。 14、 加入5ml 80％乙醇漂洗柱中的树脂 柱子真空抽干后将柱子放入用户提供的离心管中2500rpm(1300g)离心5分钟。 15、 取出柱子，真空抽干5分钟再将柱 ...

（二）碱法 1、取1.5ml培养液倒入1.5ml eppendorf管中4℃下12000g离心30秒。 2、弃上清将管倒置于卫生纸上数分钟使液体流尽。 3、菌体沉淀重悬浮于100μl溶液Ⅰ中(需剧烈振荡)室温下放置5-10分钟。 4、加入新配制的溶液Ⅱ200μl 盖紧管口，快速温和颠倒eppendorf管数次以混匀内容物(千万不要振荡)冰浴5分钟。 5、加入150μl预冷 ...

Protocol for Isolation of Genomic DNA from Mammalian Cells 1.Trypsinize harvest and resuspend cells at 107 / ml in 10 mM Tris-HCl (pH 8.0) 10 mM EDTA. 2.Add SDS and Proteinase K to a final concentrat ...

alimama_pid="mm_10043573_137377_1822161"; alimama_titlecolor="FF9900"; alimama_descolor ="552c55"; alimama_bgcolor="f7f7f7"; alimama_bordercolor="f7f7f7"; alimama_linkcolor="552c55"; ...

alimama_pid="mm_10043573_137377_1822161"; alimama_titlecolor="FF9900"; alimama_descolor ="552c55"; alimama_bgcolor="f7f7f7"; alimama_bordercolor="f7f7f7"; alimama_linkcolor="552c55"; ...

alimama_pid="mm_10043573_137377_1822161"; alimama_titlecolor="FF9900"; alimama_descolor ="552c55"; alimama_bgcolor="f7f7f7"; alimama_bordercolor="f7f7f7"; alimama_linkcolor="552c55"; ...

This a brief protocol for plasmid mini preparation.

alimama_pid="mm_10043573_137377_1822161"; alimama_titlecolor="FF9900"; alimama_descolor ="552c55"; alimama_bgcolor="f7f7f7"; alimama_bordercolor="f7f7f7"; alimama_linkcolor="552c55"; ...

alimama_pid="mm_10043573_137377_1822161"; alimama_titlecolor="FF9900"; alimama_descolor ="552c55"; alimama_bgcolor="f7f7f7"; alimama_bordercolor="f7f ...

alimama_pid="mm_10043573_137377_1822161"; alimama_titlecolor="FF9900"; alimama_descolor ="552c55"; alimama_bgcolor="f7f7f7"; alimama_bordercolor="f7f7f7"; alimama_linkcolor="552c55"; ...

alimama_pid="mm_10043573_137377_1822161"; alimama_titlecolor="FF9900"; alimama_descolor ="552c55"; alimama_bgcolor="f7f7f7"; alimama_bordercolor="f7f7f7"; alimama_linkcolor="552c5 ...

alimama_pid="mm_10043573_137377_1822161"; alimama_titlecolor="FF9900"; alimama_descolor ="552c55"; alimama_bgcolor="f7f7f7"; alimama_bordercolor="f7f ...