EXPERIMENTAL BIOLOGY BASIC CORE SKILLS LABORATORY MANUAL

This unit aims to acquire the basic core skills necessary for functioning as a laboratory scientist through various experiments.
Students will build a portfolio of competencies during four Core Skills practical sessions, and further skills through Course Specific practicals unique to their course path.
Emphasis on the practical aspect of biology, with a focus on making the unit enjoyable and enlightening.
Virtual interactive simulations are available on the Experimental Biology Moodle page to enhance skill development and understanding.
Assessment methods include:
- Formative assessment (non-contributory to final unit mark):
- Oral feedback from lecturers and demonstrators during practical lessons.
- Feedback on the laboratory report for the 2nd Core Skills practical (Spectroscopy and Standard Curve).
- Summative assessment (contributes directly to final unit mark):
- PASS/FAIL Practical Examination on Core Basic Skills.
- 50% of the unit mark based on a laboratory report for one Course Specific Practical.
- 50% based on a statistics MCQ test.

A good lab book is essential for every scientist and should be clear, well-annotated, and easily navigable.
Format to maintain a good lab book:
- Write "CONTENTS" at the top of the first page; include a list of experiments with page numbers.
- Label pages numerically.
- Record the experiment name, date, and brief description of aims at the beginning of each experiment.
- Document the procedure meticulously, noting any deviations.
- Tabulate or label results clearly and paste them into the lab book.
- When reaching the end of a page, note "continued on page…" and follow up on the next page with the preceding header.
- Reserve the last two pages for an INDEX where protocols performed are noted with corresponding page numbers.
Prioritize orderliness and ensure the cleanliness of your workspace as it reflects your lab book’s structure.

Each practical includes a COSHH (Committee on Substances Hazardous to Health) form that outlines risks associated with the experiment.
Legal requirement to read, sign, and incorporate the COSHH form into the lab book before starting any experiment.
Main laboratory risks include:
- Poisoning: Avoid ingesting hazardous materials; no eating/drinking in labs. Emphasize proper disposal of toxic substances.
- Broken Glass: Wear safety goggles, use disposal bins designated for broken glass, never work with chipped or broken glass.
- Electrocution: Be cautious when using electrical devices with aqueous solutions.
General precautions:
- Pay attention to instructions from lecturers/demonstrators.
- Use appropriate personal protective equipment (PPE).
- Always ask for guidance if uncertain about procedures.
- Maintain awareness of surroundings and potential hazards.

Accurately prepared solutions and buffers are fundamental to scientific experiments.
Skills in transferring small volumes using pipettes are essential.

Practice pipetting skills by dispensing water onto a balance and preparing solutions.
Calculate the appropriate amounts needed for various laboratory reagents:
- Tris base: 2-Amino-2-(hydroxymethyl)-1,3-propanediol, Molecular weight (Mw) = 121.14 g/mol.
- EDTA (sodium salt): Disodium ethylenediaminetetraacetic acid, Mw = 372.24 g/mol.
- Sucrose: β-D-Fructofuranosyl-α-D-glucopyranoside, Mw = 342.30 g/mol.

Equations:

Pipette Training: Familiarize with different pipettes: P20, P200, P1000, and P5000.
- Execute pipetting to measure specified volumes and ensure accuracy.
Solution Preparation:
- Solution (1): 200 ml of 200 mM Tris-HCl at pH 7.5.
- Solution (2): 200 ml of 20 mM EDTA at pH 8.0.
- Solution (3): 20 ml of 40% (w/v) sucrose.
Preparation Instructions:
- Weigh out chemicals accurately, dissolve in distilled water, adjust pH as required, and label prepared solutions.

Calibration of assay systems is critical in quantitative analysis, linking independent and dependent variables through standard curves.

Construct a fluorescence-based standard curve with methylene blue solutions for measurement purposes.

Prepare Methylene Blue Working Solution:
- Dilute original stock to create specific concentrations.
- Utilize C1V1 = C2V2 for calculations.
Standard Curve Preparation:
- Measure absorbance at 600 nm for all standard solutions.
- Document absorbance of unknown sample (Reagent A).
Graphing: Use absorbance and concentration values to create a standard curve and derive unknown concentrations based on the curve.

Utilize light microscopes (compound and stereo) for examining biological specimens.

Set up equipment for viewing various organisms through both types of microscopes.
Identify components and operate according to directions, ensuring to calibrate appropriately for focus and clarity.
Document observed specimens by making biological drawings with titles, scales, and descriptions.

PCR amplifies specific DNA sequences, utilizing primers complementary to structural DNA.

PCR Setup:
- Combine necessary materials in designated tubes while preventing contamination.
- Include specific cycle conditions for optimal amplification.
Gel Electrophoresis Analysis:
- Separate PCR products by size using agarose gel, visualizing with ethidium bromide under UV light.

Evaluate results based on band patterns and sizes in comparison to molecular markers.

Title: Clear and concise.
Introduction: Context of the experiment, background literature, and specific aims.
Materials and Methods: Detailed and replicable descriptions of procedures taken (past tense).
Results: Objective representation of data, presented with necessary annotations.
Analysis, Discussion, and Conclusion: Critical evaluation of results, relevance to hypotheses, and citations to previous studies.

Attendance at the scheduled practical examination is mandatory, covering key skills acquired during the unit.
Skills assessed include pipetting, gel loading, spectroscopy methods, microscopy skills, and record keeping.
Failure to pass this exam will result in failing the module.