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This chapter offers an inclusive overview of crucial molecular biology techniques, with a primary focus on gene cloning and its critical role in genomics, diagnostics, therapy, and biotechnology. It covers the ancient background, key enzymes, host systems, and types of cloning vectors, including plasmids, BACs, and YACs. The construction and screening of genomic and cDNA libraries are outlined in detail. Techniques for DNA amplification, such as PCR and RT-PCR, are discussed along with their applications in gene expression studies. The chapter also reviews genome mapping methods RFLP, RAPD, AFLP, and FISH and introduces MALDI-TOF mass spectrometry as a powerful tool in proteomics. It explores genome expression profiling through microarrays and ESTs, followed by key DNA sequencing techniques, from Sanger to next-generation sequencing. It concludes with advanced gene regulation strategies, focusing on RNA interference (RNAi) and the CRISPR-Cas system, emphasizing their transformative impact on genetic research, medicine, and biotechnology.

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Modern Molecular Biology: Tools, Applications, and the Power of Gene Cloning

  • 章节

  • 发表日期：

    2026年

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  • Sanjeev Kumar,
  • Lal Chuangsangi,
  • Girish Chandra Mohanta,
  • Satish Kumar Pandey

摘要

This chapter offers an inclusive overview of crucial molecular biology techniques, with a primary focus on gene cloning and its critical role in genomics, diagnostics, therapy, and biotechnology. It covers the ancient background, key enzymes, host systems, and types of cloning vectors, including plasmids, BACs, and YACs. The construction and screening of genomic and cDNA libraries are outlined in detail. Techniques for DNA amplification, such as PCR and RT-PCR, are discussed along with their applications in gene expression studies. The chapter also reviews genome mapping methods RFLP, RAPD, AFLP, and FISH and introduces MALDI-TOF mass spectrometry as a powerful tool in proteomics. It explores genome expression profiling through microarrays and ESTs, followed by key DNA sequencing techniques, from Sanger to next-generation sequencing. It concludes with advanced gene regulation strategies, focusing on RNA interference (RNAi) and the CRISPR-Cas system, emphasizing their transformative impact on genetic research, medicine, and biotechnology.