Lysate-based Systems
Lysate-based Systems - Related Videos
04:08
Purification of Bacteriophages from Bacterial Lysate
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2026
Source: Pett, N., et al. T4 Bacteriophage and E. coli Interaction in the Murine Intestine: A Prototypical Model for Studying Host-Bacteriophage Dynamics In Vivo. J. Vis. Exp. (2024)This video demonstrates the purification of bacteriophages from bacterial cell lysates using sequential centrifugation, membrane filtration, chloroform extraction, centrifugal filtration, and buffer exchange to obtain a purified phage preparation.
03:36
Preparation of Bacteriophage Lysate for Generalized Transduction
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2026
Source: Saragliadis, A., et al. Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes. J. Vis. Exp.(2018)This video demonstrates preparation of a bacteriophage lysate containing transducing particles. A donor strain is infected, plaques form, and the soft agar overlay is processed with chloroform to yield a clarified lysate for downstream analysis.
08:33
Cell-Free In Vitro mRNA Translation In Drosophila Lysates
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2026
Here, we provide details on how to prepare lysates and perform in vitro translation using Drosophila embryos.
14:42
Liquid Chromatography Coupled to Refractive Index or Mass Spectrometric Detection for Metabolite Profiling in Lysate-based Cell-free Systems
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Cited by 6 •
2021
The protocols describe high-performance liquid chromatography methods coupled to refractive index or mass spectrometric detection for studying metabolic reactions in complex lysate-based cell-free systems.
06:25
Electrochemiluminescence Assay for Quantification of Target Protein Levels in Brain Lysate
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2025
In this video, we demonstrate the electrochemiluminescence immunoassay for the quantitation of target protein levels in different samples. This assay is based on the luminescence produced in the electrochemical reaction. The intensity of light emitted is proportional to the quantity of the target protein in the sample.
FAQs
Cell disruption releases enzymes, ribosomes, nucleic acids, and other molecular components into an extract. These retained elements provide the machinery needed for biochemical activity outside intact cells. Their combined presence allows researchers to support transcription, translation, and metabolic conversion when appropriate DNA, RNA, substrates, or cofactors are supplied.
Supplied inputs act as programmable or reactive materials for the extract. DNA and RNA can direct nucleic-acid and protein-production processes, while substrates and cofactors support metabolic conversion. Changing these inputs gives researchers a way to drive different biochemical activities without relying on cellular growth or survival.
Removing the requirements for cellular growth and survival makes the reaction environment more controllable. Researchers can focus on the activity of released molecular machinery and introduce selected inputs directly. This separation supports rapid testing of genetic circuits, biosensors, and metabolic pathways while avoiding the need to maintain an intact organism during each experiment.
An intact cell coordinates biochemical production with growth and survival, whereas a lysate-based system separates those functions into an extract. This distinction gives researchers direct control over supplied DNA, RNA, substrates, and cofactors. As a result, the platform provides a more tractable setting for examining molecular mechanisms and testing engineered functions.
The workflow begins by disrupting cells to release their biological machinery and collecting the resulting extract. Researchers then supply DNA, RNA, substrates, or cofactors according to the intended reaction. The prepared mixture can support transcription, translation, or metabolic conversion, creating a controlled platform for evaluating a designed biochemical function.
These systems can support rapid prototyping of genetic circuits, biosensors, and engineered metabolic pathways. They also provide tractable models for studying molecular mechanisms because the relevant machinery operates in an extract rather than within a growing organism. The resulting flexibility makes them useful for testing biological designs and biochemical activities.
Lysate-based platforms can advance distributed biomanufacturing by enabling biological production outside intact living organisms. Their programmable extracts can receive selected DNA, RNA, substrates, or cofactors and support biochemical processes such as translation or metabolic conversion. This flexibility is relevant when production requires adaptable biological systems that can be configured for different outputs.