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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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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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 ...

The standard solution typically used for both pre-hybridisation and hybridisation is based on that given in Maniatis et al.(1982)with both Denhart's solution and heterologous DNA being replaced by hep ...

Non-radioactive Probes I. Via random hexamers 1. Solutions:10X hexa nt miundefined: 500 mM Tris-Cl pH 7.2100 mM MgCl21 mM dithioerythritol (DTE)2 mg/ml BSA62.5 A260 units/ml (1.56 mg/ml) random hexanucleotid ...

RNA/Zeta Probe Dot Blotting protocol1)Make up RNA (up to 20μg)dissolved in sterile H2 OTE or 0.5% SDS to 500μl with ice-cold sterile 10mM NaOH1mM EDTA and apply it to Zeta Probe membraneheld in ...

1. Depurination of DNA fragments: Wash gel in 0.25 M HCl 5' (small gels) 7' (large gels)2. Denaturation: Wash gel in (0.5 M NaOH 1.5 M NaCl) for 20' (small) to 30' (large).3. Neutralization: Wash in ( ...

About 1/2 to 1 cm of tail should be cut from properly marked mice (using toe or ear clipping etc) about the age of 2-3 weeks. Place these tails into marked 1.5 ml Eppindorf tubes for processing; they ...

For use with GibcoBRL Random Primer DNA labeling system. Objective: To produce radioactively labeled DNA strands for the detection of target DNA or RNA sequences in various applications including Sout ...

1) 取10μl待测DNA，于一定浓度的琼脂糖凝胶上进行电泳。(20mL，1.0%凝胶)2) 凝胶用溴化乙锭染色，切掉凝胶四周多余部分，并在凝胶的一角作一记号， 拍照记录电泳结果(拍照时， 凝胶旁放一尺子)。3) 杂交用胶的制备 (做以下步骤两组合一)A. 将凝胶浸没入30mL 0.25mol/L HCl溶液中15min，使DNA脱嘌呤。B.用蒸馏水短暂洗凝胶2次。C.将凝胶浸没入30mL变 ...

一 酶切和电泳在200 μl 微量离心管中加入：25 μl DNA样品(约10μg)，3μl 限制性内切酶（MBI，10 U/ μl）5 μl 相应的10×buffer，补水到50μl。然后加一滴矿物油覆盖 稍微离心后放于37℃水浴8-12小时。酶切完后，取5μl 酶切DNA样品于0.8%的琼脂糖凝胶上检测酶是否充分。如果酶切充分，灌制0.8%的ag ...