Method for selection of DNA molecules with a desired nucleotide sequence
Andrey Pichugin, Markozashvili D, Nesterov A, Bardin A, Kim E, Langin S, Maharova M, Barinov A, and Vassetzky Y
Peter the Great St. Petersburg Polytechnic University, St. Petersburg, Russia
UVSQ-Inserm U1179 - End-icap, Handicap Neuromusculaire - Physiopathologie, Biothérapie et Pharmacologie appliquées, UFR des sciences de la santé
: J Pharm Sci Emerg Drugs
Abstract
New methods to produce DNA molecules with the desired nucleotide sequence are required in the field of synthetic biology. We developed an NGS-related method for selection of DNA molecules. Like in Next Generation Sequencing methods, the dNTPs are added in a sequential manner to the immobilized primed templates in order to perform DNA-polymerase-dependent elongation of primers. In our method, the templates may be released from the solid support into a supernatant by a stranddisplacement activity of DNA-polymerase. The error-free templates are obtained from the supernatant and the non-desired templates with mutations remain immobilized on the solid support. The reaction conditions have been determined and fully automated.
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