Chapter 1 Strategies for Novel Research Projects ............................. 1 Chapter 2 Rapid Isolation of Specific cDNAs or Genes by PCR ................. 15 Chapter 3 Construction and Screening of Subtracted and Complete Expression cDNA Libraries ......................................... 31 Chapter 4 Subcloning of Genes or DNA Fragments .............................. 73 Chapter 5 Nonisotopic and Isotopic DNA or RNA Sequencing .................... 99 Chapter 6 Information Superhighway and Computer Databases of Nucleic Acids and Proteins ............................................... 139 Chapter 7 Characterization of DNA or Genes by Southern Blot Hybridization .................................................... 165 Chapter 8 Gene Overexpression by Sense RNA in Mammalian Systems ............ 193 Chapter 9 Gene Underexpression in Cultured Cells and Animals by Antisense DNA and RNA Strategies ................................. 223 Chapter 10 Analysis of Gene Expression at the Functional Genomic Level ..... 247 Chapter 11 Analysis of Gene Expression at the Proteomic Level .............. 265 Chapter 12 Analysis of Cellular DNA or Abundance of mRNA by Radioactivity in Situ Hybridization (RISH) .................................... 287 Chapter 13 Localization of DNA or Abundance of mRNA by Fluorescence in Situ Hybridization (FISH) .................................... 311 Chapter 14 In Situ PCR Hybridization of Low Copy Genes and in Situ RT-PCR Detection of Low Abundance mRNAs ......................... 323 Chapter 15 Isolation and Characterization of Genes from Genomic DNA Libraries ....................................................... 341 Chapter 16 Culture of Mouse Embryonic Stem Cells as a Model Mammalian Cell Line for Gene Expression ................................... 365 Chapter 17 New Strategies for Gene Knockout ................................ 375 Chapter 18 Large-Scale Expression and Purification of Recombinant Proteins in Cultured Cells ...................................... 409 Chapter 19 Quantitative Analysis of Functional Genome by Current Real-Time RT-PCR ................................................ 445 Chapter 20 High-Throughput Analysis of Gene Expression by Cutting-Edge Technology — DNA Microarrays (Gene Chips) ....................... 461 Chapter 21 Construction and Screening of Human Antibody Libraries: Using State-of-the-Art Technology — Phage Display ..................... 471 Chapter 22 Down-Regulation of Gene Expression in Mammalian Systems via Current siRNA Technology .................................... 493 Index ...................................................................... 509