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The preferred method of de-phosphorylation uses the buffer system described by Pharmacia for most of their restriction endonucleases. You will need: 10 x OPA+ Buffer (100mM Tris.acetate pH 7.5 100mM ...

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The gel shift or electrophoretic mobility shift assay provides a simple and rapid method for detecting DNA -binding proteins. This method has been used widely in the study of sequence-specific DNA -bi ...

Restriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restriction enzyme in its respective buffer as recommended by the supplier and at ...

It is best to do a test ligation without insert if the background is low it would not be necessary to screen colonies for insert. 1.colony PCR 1) Pick a colony from plate and suspend the colony in 10 ...

简介 优点：操作简单方便 缺陷：不能像EMSA或footprinting一样区分不同类型复合物 原理 双链 DNA 能够通过硝化纤维膜 (NC)，但是 DNA-protein 复合物不能通过NC filter. 通过定量分析留在滤膜上的复合物，可以判断二者的结合常数。 基本方法 Lable DNA Mix DNA and protein to form complex Pass th ...

Sample Preparation Cheek cells are obtained by rinsing the mouth with 25mls of any commercial mouth wash solution available for about 30sec the first thing in the morning. It is important not to brus ...

This is a rapid method for chemical DNA sequencing which is commonly used as ladder for footprinting reactions or for sequencing of short DNA oligonucleotides. Reference: Bencini et al. (1984) Biotech ...

Source: Protocol Online Abstract: Modifying DNA using sodium bisulfite to convert unmethylated cytosines to uracils and subsequently detect methylated cytosines using methylation specific PCR (MSP) te ...

Miniprep of BAC DNA 1.Inoculate 4 - 8 tubes of 3ml/tube LB with 15μg/ml chloramphenicol with BAC clone and grow oveernight. Miniprep with AutoGen 740 according to the protocol described in our Web ...

The following protocol is designed for subcloning inserts (I) from one vector into another vector (V). The inserts can be anywhere from 30 bp to 8 kb (possibly higher). Perform restriction diges ...

Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute http://www.fhcrc.org/science/labs/hahn/methods/mol_bio_meth/Big%20Dye%20Protocol.pdf ...

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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 ...

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 So ...

METHOD: 1.Isolate chromosomal DNA by using the CTAB protocols in current Pr M R. 2.Make a CsCl preparation of pLAFr3 (or other appropriate cosmid). 3.Cut pLAFr with BamHI phenol ext. + ETOH preparatio ...