Molecular cloning of DNA from a sorted human minichromosome.
A human supernumerary minichromosome (MC), found in a newborn baby and sorted on a fluorescence-activated cell sorter (FACS-440) has been previously described [Ferretti et al., Cytotechnology 1 (1987) 7-12]. We report here on the construction of a library of EcoRI fragments in the phage lambda gtWES.lambda B', starting from 7.5 ng of MC DNA, and describe the isolation of single-copy DNA clones from the library in a two-step procedure. We employed in situ hybridization to unambiguously select the clones specific for the MC, and used three of these clones to demonstrate that it originated from chromosome 9.
Ferretti
L
L
Istituto per la Difesa e la Valorizzazione del Germoplasma Animale, CNR, Milan, Italy.
Raimondi
E
E
Gamberi
C
C
Young
B D
BD
De Carli
L
L
Sgaramella
V
V
D002899:Chromosomes, Human, Pair 9; D003001:Cloning, Molecular; D015342:DNA Probes; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D015723:Gene Library; D006801:Humans; D015151:Immunoblotting
D015342:DNA Probes; D004274:DNA, Recombinant