Three yeast artificial chromosome (YAC) libraries were constructed using two human cell lines and the pYAC-RC vector. The main differences from the previously described methods were: i) genomic DNA was digested in low melting point (LMP) agarose blocks with the rare cutting enzyme ClaI; ii) DNA was ligated in melted LMP agarose after agarase treatment; iii) spheroplast regeneration plating was done in calcium alginate thin layer. In addition, a panel of PCR primers was used to identify quickly the presence in the libraries of repetitive and single copy human DNA sequences.
Titolo: | Simplified construction and characterization of yeast artificial chromosome libraries |
Autori interni: | |
Data di pubblicazione: | 1992 |
Rivista: | |
Handle: | http://hdl.handle.net/20.500.11769/335194 |
Appare nelle tipologie: | 1.1 Articolo in rivista |
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