[47] Using bacteriophage P1 system to clone high molecular weight genomic DNA

摘要: Publisher Summary This chapter describes the use of bacteriophage p1 system to clone high molecular weight genomic DNA. The phage P1 can with an efficiency that makes it practical generate mammalian libraries reasonable ease (greater than 105 clones be generated per microgram vector and insert) several micrograms isolated insert DNA produced from 109 cells by standard plasmid isolation procedures. A partial 50,000-member human library a complete Drosophila have been constructed. Complete are being produced. Bacteriophage packages its processive headful mechanism. substrate for this reaction is concatemer rolling circle replication consists tandemly repeated units genome arranged in head-to-tail configuration. entire screened one agarose gel polymerase chain (PCR) or southern blot analysis secondary pools. If pool positive, made 10 primary pools represent analyzed same way. generates positive signal processed colony hybridization select individual desired insert.