ABSTRACT
Filled-In DNA Fragments .......................................................................... 106 6.2.2 Screening of the Genomic DNA Library ................................................................. 107
6.2.2.1 Screening of the Genomic Library with a 32P-Labeled Probe ................... 107 6.2.2.2 Screening of a Genomic Library Using a Nonradioactive Probe .............. 110
6.2.3 Construction of Partial Genomic Libraries .............................................................. 110 6.2.3.1 Southern Blotting of Genomic DNA and Identi cation
of Desired Fragments ................................................................................. 111 6.2.3.2 Isolation of Desired Fragments .................................................................. 111 6.2.3.3 Ligation of Linkers .................................................................................... 111 6.2.3.4 Construction and Screening of the Library ............................................... 111
6.2.4 Restriction Mapping of Positive Recombinant Bacteriophage DNA Clones ........... 111 6.2.5 Subcloning of the DNA Fragment of Interest ........................................................... 113
6.2.5.1 Restriction Enzyme Digestion of Vector and DNA Insert for Subcloning .................................................................................... 114
6.2.5.2 Ligation of Plasmid Vector and Insert DNA .............................................. 116 6.2.5.3 E. coli Transformation ............................................................................... 117 6.2.5.4 Transformation Using Chemically Competent Cells ................................. 118 6.2.5.5 Transformation by Electroporation ............................................................ 119 6.2.5.6 Selection of Transformants Containing Recombinant Plasmids ............... 120
6.2.6 Reagents Needed ...................................................................................................... 120 6.3 YAC Libraries .......................................................................................................................124
