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The RPA-coated ssDNA generated during the extensive end resection mediated through HR nucleases is processed via several HR subpathways, depending on the homologous DNA availability (reviewed in San Filippo, Sung and Klein 2008).
Consistently, defects of sae2 null cells in DSB processing after exposure to ionizing radiation are suppressed by inactivation of YKU70, and this suppression requires both nucleases mediating the extensive end resection, Exo1 and Sgs1 Dna2 (Mimitou and Symington 2010).
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The extensive end-stage screening and sequencing of bacterial clones made this protocol, as it stood, impractical.
In contrast, HR is a high-fidelity mechanism, using a chromosomal homologue as a template for the repair, and involves the extensive end-resection of the break.
The rate of extensive end resection is ∼4 kb/hr and can remove up to 50 kb of DNA if there is no donor sequence to template repair of the DSB (Zhu et al. 2008).
The y− w− progeny results from loss of the y + coding sequence by extensive end resection (>4.4 kbp), followed by end-joining (incomplete HR) or extensive end resection of >7.2 kbp, followed by SSA of the direct repeat sequence.
An essential step during HR-mediated repair is the extensive 5′ 3′ end resection of the DSB.
The inability of spn-A mutants to strand invade resulted in extensive end resection of >7.2 kb, followed by SSA.
The story comprises 241 pages of the 271-page book, and is supplemented with a complete scholarly apparatus: a foreword, bibliography, index, map section and extensive end notes.
Alan Moore's use of extensive end notes in his and Eddie Campbell's From Hell, another fictional reconstruction of a historical event, influenced Brown's appendices.
In checkpoint deficient strains extensive end degradation up to and past a centromere could provide for an alternative mechanism to initiate secondary rearrangements of dicentric GCRs in addition to other breakage mechanisms including mechanical breakage or inverted repeat-induced DSBs.
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