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A Novel General Chemistry Experiment for Freshmen: Transgenic Soybean Detection Based on Microfluidic Molecular Fluorescence Spectroscopy Analysis

A Novel General Chemistry Experiment for Freshmen: Transgenic Soybean Detection Based on Microfluidic Molecular Fluorescence Spectroscopy Analysis
Learning Objets
Summary
This educational resource introduces a microfluidic-based molecular fluorescence spectroscopy experiment designed for freshmen enrolled in General Chemistry courses. The experiment focuses on the detection of transgenic soybeans by analyzing nucleic acids using a centrifugal microfluidic platform combined with loop-mediated isothermal amplification (LAMP). It serves as an interdisciplinary teaching module, integrating principles of molecular fluorescence spectroscopy, enzyme reaction kinetics, and microfluidic technology to enhance students' understanding of chemistry in real-world applications. The course has been successfully implemented at over 30 universities in China, with high student engagement and positive feedback. The experiment fosters hands-on skills, critical thinking, and interest in analytical chemistry and biotechnology.

Authors/Contributors: Pintao Li, Min Gu, Ghazala Ashraf, Huiwen Xiong, Fei Cun, Xuting Chen, Jilie Kong, and Xueen Fang.

Citation: Li, P., Gu, M., Ashraf, G., Xiong, H., Cun, F., Chen, X., Kong, J., & Fang, X. (2024). A Novel General Chemistry Experiment for Freshmen: Transgenic Soybean Detection Based on Microfluidic Molecular Fluorescence Spectroscopy Analysis. Journal of Chemical Education, 101, 4353-4359. https://doi.org/10.1021/acs.jchemed.4c00668
Keywords
Microfluidics
Nucleic Acids
Molecular Fluorescence Spectroscopy
Transgenic soybean
General Chemistry Laboratory

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Moderation state
Published
Object Type
Laboratory experiment
Journal articles
Audience
High School (Secondary School)
Introductory Undergraduate
Published on
Green Chemistry Principles
Waste Prevention
Atom Economy
Safer Solvents and Auxiliaries
Design for Energy Efficiency
Use of Renewable Feedstocks
U.N. Sustainable Development Goals (SDGs)
Quality Education
Industry, Innovation and Infrastructure
Responsible Consumption and Production
Climate Action
Life on Land
NGSS Standards, if applicable
The experiment aligns with several Next Generation Science Standards (NGSS), particularly in the areas of Physical Science (PS), Life Science (LS), and Engineering, Technology, and Applications of Science (ETS).

Applicable NGSS Standards:
High School - Physical Science (PS)
HS-PS1-1: Use the periodic table as a model to predict the relative properties of elements based on the patterns of electrons in the outermost energy level of atoms.
Relevance: Understanding fluorescence spectroscopy involves knowledge of atomic structure, electron excitation, and emission spectra.

HS-PS1-3: Plan and conduct an investigation to gather evidence to compare the structure of substances before and after a chemical reaction.
Relevance: The experiment involves nucleic acid amplification and fluorescence emission, demonstrating chemical reactions in biological systems.

High School - Life Science (LS)
HS-LS1-1: Construct an explanation based on evidence for how the structure of DNA determines the structure of proteins, which carry out essential functions of life through systems of specialized cells.
Relevance: The experiment involves DNA extraction, amplification, and detection, reinforcing concepts of genetic structure and function.

HS-LS3-1: Ask questions to clarify relationships about the role of DNA and chromosomes in coding the instructions for characteristic traits.
Relevance: The study of transgenic soybeans directly relates to genetic modification and inheritance patterns.

Engineering, Technology, and Applications of Science (ETS)
HS-ETS1-2: Design a solution to a complex real-world problem by breaking it down into smaller, more manageable problems that can be solved through engineering.
Relevance: Microfluidic technology in this experiment serves as a real-world engineering application in biotechnology and analytical chemistry.
This experiment provides hands-on, inquiry-based learning that aligns with NGSS practices such as developing models, conducting investigations, analyzing data, and constructing explanations based on evidence.
Learning Goals/Student Objectives
By completing this experiment, students will be able to:
1. Understand Microfluidic Technology & DNA Analysis:
a. Explain the principles of microfluidics and its applications in chemical and biological analysis.
b. Describe the fundamentals of nucleic acid amplification and molecular fluorescence spectroscopy.

2. Apply Chemistry Concepts to Real-World Scenarios:
a. Understand the chemical principles behind nucleic acid detection, including enzyme kinetics, fluorescence emission, and molecular interactions.
b. Explain the role of DNA amplification in biotechnology, including applications in genetic screening and transgenic crop detection.

3. Develop Hands-On Laboratory Skills:
a. Extract DNA from soybean samples using a high-temperature alkali boiling method.
b. Perform microfluidic-based fluorescence analysis and interpret results.
c. Construct a standard curve and use it to quantify nucleic acid concentrations.

4. Enhance Data Analysis & Critical Thinking Abilities:
a. Analyze experimental data, including fluorescence intensity curves and amplification efficiency.
b. Evaluate sources of error and troubleshoot issues in experimental procedures.
Interpret results to determine whether a soybean sample is transgenic.

5. Encourage Multidisciplinary Thinking:
a. Recognize how chemistry integrates with biology, physics, and engineering in real-world applications.
b. Discuss the broader implications of genetic modification in agriculture, medicine, and environmental science.

6. Improve Scientific Communication & Collaboration:
a. Work collaboratively in a laboratory setting, following safety protocols and best practices.
b. Communicate findings through written lab reports and presentations, demonstrating a clear understanding of experimental results.
Common pedagogies covered
Blended learning
Collaborative/cooperative learning
Context-based learning
Hands-on learning

Submitted by

Kris Weigal
Safety Precautions, Hazards, and Risk Assessment
Chemical Hazards
Sodium Hydroxide (NaOH) (Used in DNA extraction)

Hazard: Corrosive to skin and eyes; can cause burns upon contact.
Mitigation:
Always wear gloves, goggles, and a lab coat.
Handle under a fume hood or well-ventilated area.
If contact occurs, rinse immediately with plenty of water and seek medical assistance.
Fluorescent Dyes (Used in nucleic acid detection)

Hazard: Some fluorescent dyes may be toxic, mutagenic, or carcinogenic upon prolonged exposure.
Mitigation:
Handle with gloves and avoid inhalation or skin contact.
Dispose of used dyes according to hazardous waste regulations.
Equipment Hazards
Centrifuge (Used for microfluidic chip operation)

Hazard: Incorrect usage can cause mechanical injury or sample spillage due to improper balance.
Mitigation:
Ensure samples are properly balanced before spinning.
Do not open the lid until the rotor has completely stopped.
Heating Plate (Used in DNA extraction)

Hazard: Can cause burns or fire risk if misused.
Mitigation:
Always use heat-resistant gloves when handling.
Keep flammable materials away from the heating area.
Biological Hazards
Nucleic Acid Samples (Extracted from Soybeans)
Hazard: Although not infectious, improper handling can lead to cross-contamination.
Mitigation:
Keep workspaces clean and decontaminated.
Use separate pipettes for different sample types.

Risk Assessment and Classification (Hazard, Risk Level and Control Measures)

Sodium Hydroxide (NaOH)
Medium
Use PPE, work in a fume hood, rinse spills immediately

Fluorescent Dyes
Medium
Wear gloves, minimize exposure, proper disposal

Centrifuge Operation
Medium
Balance samples, do not open until stopped

Heating Plate
Medium
Use heat-resistant gloves, keep flammables away

Nucleic Acid Handling
Low
Avoid cross-contamination, use proper pipetting techniques
Link to external
https://pubs.acs.org/doi/10.1021/acs.jchemed.4c00668