Desalting DNA by Drop Dialysis Increases Library Size upon Transformation

It is often desirable to obtain gene libraries with the greatest possible number of variants. We tested two different methods for desalting the products of library ligation reactions (silica-based microcolumns and drop dialysis), and examined their effects on final library size. For both intramolecu...

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Veröffentlicht in:	Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2013, Vol.77 (2), p.402-404
Hauptverfasser:	SARASWAT, Mayank, GRAND, Ralph. S., PATRICK, Wayne M.
Format:	Artikel
Sprache:	eng
Schlagworte:	Adsorption Cloning, Molecular - methods Colony Count, Microbial directed evolution DNA - genetics DNA - isolation & purification drop dialysis Electroporation Escherichia coli - genetics Gene Library Genetic Vectors - isolation & purification library construction ligation microcolumn Microdialysis - methods Microdialysis - standards Salts - chemistry Transformation, Bacterial
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Zusammenfassung:	It is often desirable to obtain gene libraries with the greatest possible number of variants. We tested two different methods for desalting the products of library ligation reactions (silica-based microcolumns and drop dialysis), and examined their effects on final library size. For both intramolecular and intermolecular ligation, desalting by drop dialysis yielded approximately 3-5 times more transformants than microcolumn purification.
ISSN:	0916-8451 1347-6947
DOI:	10.1271/bbb.120767