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The integration of nanotechnology in medicine has had a tremendous impact in the past few decades. The discovery of synthesis of nanomaterials (NMs) and their functions as versatile tools promoted various applications in nano-biotechnology and nanomedicine. Although the physical and chemical methods are still considered as commonly used methods, they introduce several drawbacks such as the use of toxic chemicals (solvent, reducing, and capping agents) and poor control of size, size distribution, and morphology, respectively. Additionally, the NMs synthesized in organic solvents and hydrophobic surfactants rapidly aggregate in aqueous solutions or under physiologic conditions, limiting their applications in medicine. Many of the phase-transfer strategies were developed and applied for the transfer of NMs into aqueous solutions. Although great efforts have been put into phase transfers, they mostly include expensive, time-consuming, intensive labor work, multi steps, and complicated procedures.Use of plant extracts in the biological synthesis method offers stark advantages over other biomolecules (protein, enzyme, peptide, and DNA). Plant extracts have been commonly used for food, medicine, NM synthesis, and biosensing. There are many viable techniques developed for the production of plant extracts with various contents based on their simplicity, cost, and the type of extract content. In this chapter, we conduct a comparative study for extract preparation techniques, the use of extracts for metallic single and hybrid nanoparticle (NP) synthesis, and their antimicrobial properties against pathogenic and plant-based bacteria. Graphical Abstract.