The CTAB-DNA precipitation method: a common mini-scale preparation of template DNA from phagemids, phages or plasmids suitable for sequencing.
This report describes a common method of obtaining template DNA from phagemids, phages and plasmids. The strategy is based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB) for DNA precipitation. By avoiding phase separation, many manipulation steps are reduced. A time-saving modification to perform double-stranded DNA sequencing directly after alkaline-denaturation is also introduced. The protocols described here allow the researcher to obtain template DNA from a variety of initial sources, thus giving reproducible sequencing results when using T7 DNA polymerase.
Del Sal
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G
International Center of Genetic Engineering and Biotechnology, Trieste, Italy.
Manfioletti
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G
Schneider
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C
D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D001709:Biotechnology; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D011232:Chemical Precipitation; D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification*; D003902:Detergents; D005821:Genetic Techniques; D005822:Genetic Vectors; D010957:Plasmids
D002593:Cetrimonium Compounds; D004277:DNA, Single-Stranded; D003902:Detergents; D000077286:Cetrimonium