Construction and preliminary analysis of the ICRF human P1 library.
P1 clone libraries have now been established as effective complements to cosmid and yeast artificial chromosome libraries in long-range mapping projects. To allow general access to P1 clones, we have constructed human and mouse P1 libraries. Clones have been picked into microtiter plates and used to prepare high-density filter grids, providing an efficient and easy screening system. Filters are being made available to other laboratories through the Reference Library System. In this work, we have developed a reliable protocol for generating P1 clones, based on the use of pulsed-field gel electrophoresis for size selection of DNA. A 1.2x genome coverage human library has been produced using this method. A preliminary analysis of this library is described.
Francis
F
F
Genome Analysis Laboratory, Imperial Cancer Research Fund (ICRF), London, England.
Zehetner
G
G
Höglund
M
M
Lehrach
H
H
D017101:Bacteriophage P1 / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D016521:Electrophoresis, Gel, Pulsed-Field; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics*; D015894:Genome, Human; D006801:Humans; D008214:Lymphocytes; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization
D004274:DNA, Recombinant; D004247:DNA