DNA实验技术

Molecularpourous membrane tubing is used for desalting protein and DNA isolation / purification. It comes in several diameters and pore sizes (for molecular weight conversions for nucleic acids see ge ...

Purification of DNA from agarose gels is an essential method involved in the sub-cloning of DNA fragments. The following method describes a variation of the method of Vogelstein and Gillespie 1979 (Pr ...

Pellet 1.5 ml of an overnight culture at 12000 rpm in Eppendorf centrifuge at RT for 3 min. Resuspend bacterial pellet in 350 µl of STET buffer. Add 25 µl of freshly prepared solution of lysozyme (10 ...

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 immediately to preve ...

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 21) at ...

作分子生物学实验，核酸纯化、酶切、克隆算是最基本的步骤了。因为要做定向克隆，通常要作双酶切。为了省时省事儿，我们常常想方设法把两个酶放在一起切。可是事情并非总那么称心如意――两个酶经常在一起闹别扭，不是反应温度不同啦，就是Buffer不同，有时候两个酶切位点连在一起，必须先切一个再切另一个，否则就切不动――因为有的酶除了识别位点之外还需要旁边留有几个碱基，如果正好被切掉了就出问题。于是有人就到处找 ...

1、将已经电泳确定的可回收的酶切产物在合适浓度的回收用琼脂糖凝胶进行电泳。最好换用新的电泳缓冲液10 × TAE（10 ×Tris-乙酸）。2、当溴酚蓝迁移至足够距离时（至少2 cm以上），在长波紫外灯下观察，用清洗过的刀片在目的片段前切下与目的片段同长，宽度适当（一般2 cm左右）的胶块。注意：①不要忘记在胶下垫一个新的塑料手套防止污染。②小心不要将回收胶切裂，同时注意与目的带相邻的切面要尽量平 ...

This is a modification of the procedure in Short Protocols (1-411-45)1. Prepare a 50 ml liquid lysate:A. mix 2xlO8 E. coli cells with 100 ul phage (from one picked plaque in 500 ul SM) and100 ul lOmM ...

Use of oligonucleotides in various research applications requires certain basic storage and handling techniques in order to ensure trouble-free experiments. Proper storage of your oligonucleotide will ...

在分子生物学研究中，DNA 的序列分析是进一步研究和改造目的基因的基础。目前用于测序的技术主要有Sanger等（1977）发明的双脱氧链末端终止法和Maxam和 Gilbert（1977）发明的化学降解法。这二种方法在原理上差异很大，但都是根据核苷酸在某一固定的点开始，随机在某一个特定的碱基处终止，产生A，T，C，G四组不同长度的一系列核苷酸，然后在尿素变性的PAGE胶上电泳进行检测，从而获得DN ...

Inoculate 1 ml of L-broth or other rich media with cells of the strain from which genomic DNA is to be isolated. Grow this culture at 37℃ overnight on a roller.Transfer this overnight culture to a 1.7 ...

This protocol was written by Jean-Pierre Issabased on Adams et al.* Kam-Wing Jair has made some useful shortcuts that work well if you are careful.Here is .This assay can be used to measure activity i ...

DNA methylation is an epigenetic event that affects cell function by altering gene expression and refers to the covalent addition of a methyl group catalyzed by DNA methyltransferase (DNMT) to the 5-c ...

FootprintingFootprinting is a method for determining the exact DNA sequence to which a particular DNA-binding protein binds. Examples: hormone-receptor complexes that bind to their hormone response el ...

Contributed by Dr. A. GratchevSingle Nucleotide Primer Extension is a powerful method which can be used for the precise analysis of methylation in a certain position. The procedure is shown on the fig ...

1. DNA DNA template plasmid 5-20 ng 　　10x pfu DNA polymerase buffer 5.0 µl 　　25uM oligo 1 0.5 µl 　　25uM oligo 2 0.5 µl 　　10mM dNTP 1.0 µl 　　Pfu DNA polymerase (2.5 units) 1.0 µl 　　fill w/ddH2O to 50 µ ...

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The Erase-a-Base® System is designed for the rapid construction of plasmid or M13 subclones containing progressive unidirectional deletions of any inserted DNA . The system is based on the procedure d ...

Mutagenesis is a fundamentally important DNA technology which seeks to change the base sequence of DNA and test its effect on gene or DNA function. The mutagenesis can be conducted in vivo (in studies ...

Southern Analysis Procedure (for analysis of genomic DNA or ordering of clones): 1) Prepare genomic DNA (ref.p.9.22 Maniatis) from cultured cells using one T75 flask (wash two times with PBS and then ...