Yeast artificial chromosome segregation from host chromosomes with similar lengths |
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Authors: | KI Izvolsky VV Demidov NO Bukanov MD Frank-Kamenetskii |
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Affiliation: | Center for Advanced Biotechnology, Boston University, 36 Cummington Street, Boston, MA 02215, USA. |
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Abstract: | We propose a new method for segregation of yeast artificial chromosomes (YACs) from endogenous yeast chromosomes with similar lengths. The method is based on recently developed PNA-assisted rare cleavage (PARC) of genomic DNA. We apply the PARC procedure to YAC-containing samples of yeast DNA in such a way that host chromosomes, which electrophoretically comigrate with the chosen YACs, are selectively digested while YACs remain intact. These data demonstrate that a pool of appropriate PNAs can be used as an efficient tool for the PARC-based isolation of intact purified YACs directly from the host cells. |
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