The use of spun columns in the purification of double-stranded cDNA appropriate for tailing |
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| Authors: | S Pan P J Doherty |
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| Affiliation: | 1. The Wistar Institute of Anatomy and Biology, 36th Street at Spruce, Philadelphia, Pennsylvania 19104 USA;2. Department of Immunology, Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, Canada M5G 1X8 |
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| Abstract: | A reliable estimation of the efficiency of adding homopolymeric tails to double-stranded cDNA molecules by terminal deoxynucleotidyl transferase is extremely important in the construction of cDNA libraries. Appreciable differences in transformation efficiency result when the homopolymer tails to be annealed are of inadequate length. We report here that the use of a Sephadex G-50 spun column to remove oligo(dT)12-18 frequently coprecipitating in ethanol with the cDNA results in a more dependable estimation of tail lengths. |
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