Titlebook: DNA Sequencing Protocols; Hugh G. Griffin,Annette M. Griffin Book 19931st edition Springer Science+Business Media New York 1993

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向前變橢圓

M13 Cloning Vehicles,h and the two DNA strands simultaneously. For this purpose, a bacteriophage like M 13 can be used. The various viral . and . functions are critical not only for strand separation, but also to separate the single-stranded DNA from the . cell by an active transport mechanism through the intact cell wall.

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Plasmid Sequencing,additional advantage of cleaning-up the template by removing traces of low molecular weight compounds that may interfere with the sequencing reactions. The method used here to prepare plasmid is a variation on the boiled-lysis method (.) using a rapid approach avoiding phenol extraction.

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M13 Phage Growth and Single-Strand DNA Preparation,f . (.). Well-separated plaques contain cells infected with phages derived from a single transformation event and these can be picked and regrown to provide pure stocks of recombinant phage particles and DNA.

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David Muhr,Michael Affenzeller,Josef Küngoresis, that IS, during fixing, drying, and autoradiography. The methods given in this chapter deal only with the “standard“ size sequencing gels, but the principles are the same for larger gels, except that more gel mix will be needed, and different sizes of combs, spacers, and glass plates may be needed

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Arnaud Castelltort,Anne Laurent can also serve as satisfactory template DNA. It is particularly important when preparing plasmid DNA for sequencing to give the utmost care and attention to the DNA isolation and purification techniques. Most problems that occur with plasmid sequencing are related to poor quality template.

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