Medical Biochemistry I Lab: Techniques Overview

Course Information

• Course: Medical Biochemistry I
• Lab Exercise 1: Introduction to Techniques - Use of Pipetman, Sample Preparation and Dilutions
• Semester: Fall 2024
• Instructors: Andreou M & Kyriakou TC
• Institution: European University Cyprus, School of Medicine

Objectives

• Introduction: Lab characteristics/guidelines
• Part A: Use of Pipetman (Theoretical Background and Exercises)
• Part B: Sample Preparation (Main units of concentration and exercises)
• Part C: Dilutions (Dilution Formula and exercises)

COVID-19 Safety Protocol

• Be on time to avoid crowds.
• Always bring protective gear (lab coat with cuffed sleeves, mask, goggles).
• Students without all gear will not be admitted.
• Practice hand hygiene (wash hands with soap, then gloves).
• Disinfect working surfaces before and after experiments.
• Report any symptoms immediately (fever, cough, breathing difficulties).

Absences

• Mandatory participation required.
• Physical presence or synchronous participation (permission required).
• Two justified absences excused (severe health condition or emergency).
• Absences must be announced before class begins.
• Lab experiments cannot be repeated.

Preparation

• Study uploaded material.
• Print protocol/experimental methodology.
• Identify materials/equipment needed from the protocol.
• Be prepared for technically-demanding applications.
• Closed-toe shoes, lab coat, goggles, and gloves required.
• Hair must be tied back (ponytail).
• No food/drinks allowed.

Lab Notebook

• Record everything explained.
• Note protocol applications with dates.
• Calculate reagent concentrations (e.g., C1V1=C2V2).
• Evaluate results and identify areas needing adjustment.

Lab Etiquette

• Move carefully (as if in a bubble).
• Be aware of your surroundings.
• Work cooperatively as a team.
• Respect others.
• Ask questions before proceeding.
• Avoid sudden movements.
• Work quietly.

Contact Information

• Email: [email protected] & [email protected]
• Office: N30 (2nd floor - Medical Building)
• EUC account policy: Faculty emails must come through EUC account.

Materials and Equipment

• Pipetman (P1000, P200, P10)
• Pipet tips
• Distilled Water
• 1.0 M CuSO4 solution
• Cuvettes
• Balance machine
• Spectrophotometer
• Weighing boats
• Waste containers

Lab Exercise 1 Details

• Introduction to techniques: use of Pipetman.
• Sample preparation.
• Dilutions.

Pipetman Use

• For volumes under 1 mL, the Pipetman is the standard method.
• Types: P1000 (200-1000 µL), P100 (10-100 µL), P10 (0.5-10 µL)
• 1 L = 1000 mL and 1 mL = 1000 µL

Pipetman Procedure (diagram reference)

Pipette Accuracy Check

• Weigh water to check pipette accuracy.
• Density of water at room temperature is 1 g/mL, meaning each microliter should weigh 0.001 g.

Part A - Pipette Use Exercises

• Tables for calculating volume using 1000µl, and 200µl pipettes (various measurement exercises for volumes).

Part B - Sample Preparation

• Solvent and solute definitions.
• Aqueous solutions defined.
• Common concentration units (%w/w, %v/v, %w/v, Molarity).
• Concentration calculation example of mass concentration (100g solution, 5g NaOH).

Part C - Dilutions

• Dilution formulas (C1V1 = C2V2) for calculating dilutions.
• Definitions of C1 (initial/stock solution concentration), V1 (volume of stock solution), C2 (final solution concentration), V2 (final solution volume).
• Parallel dilution, and various dilution factors (e.g., 10X, 20X, 100X solutions).
• Example dilution problem for converting 10X NaOH to 1X.