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

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Andrés Iglesias,Akemi Gálvez,Marta Collanteseating the DNA with alkali. Conventionally, use of the alkali denaturation method involves neutralization of the sample by acid treatment followed by ethanol precipitation and recovery of the DNA by centrifugation. The time-consuming process of ethanol precipitation may be avoided by neutralizing th

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Springer Science+Business Media New York 1993

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M13 Phage Growth and DNA Purification Using 96 Well Microtiter Trays,of samples is tedious, however, and much time is spent opening and closing tubes and transferring tubes in and out of microcentrifuges. One hundred small volume phenol extractions and ethanol precipitations tasks even the more dedicated sequencer.

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Generation of Random Fragments by Sonication,d. A high proportion of large-scale sequencing projects use random cloning and sequencing (shotgun sequencing) to produce in excess of 95% of the data, and then employ more directed means to complete it (.–.).

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Pouring Linear and Buffer-Gradient Sequencing Gels,oresis, 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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Plasmid Sequencing, 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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