Simplified construction and characterization of yeast artificial chromosome libraries.
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.
Volinia
S
S
Instituto di Chimica Biologica, Universit, Ferrara, Italy.
Piva
R
R
Bozza
A
A
Stefani
S
S
Gandini
D
D
del Senno
L
L
D001483:Base Sequence; D002460:Cell Line; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA; D015723:Gene Library; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data
D004247:DNA