Introduction to Equipment Biotechnology Fall 2012

Lesson 1: Essential Questions What are the most common pieces of biotechnology laboratory equipment? What is aseptic technique? Why is it important? What happens when we do not practice it?

Lesson 1: Learning Objectives By the end of today, you should be familiar with the most common pieces of lab equipment You should know what each piece of equipment is and what it is used for In addition, you should know what aseptic technique is and why it is important in the biotechnology laboratory

Measurements In the science laboratory we use the metric system To measure volume= liter To measure mass or weight= gram To measure length= meter However, Biotechnology often works on the milli and micro-scales (103 and 106)

Volume To measure large volumes (greater than 25 ml) you would use a graduated cylinder They come in several sizes They are very accurate Measure to meniscus

Volume Larger volumes (greater than 25 ml) should be made in beakers and flasks These should not be used to measure, as they are not very accurate

Volumes Small amounts are usually measured with pipets There are two types Glass pipets (2-25 ml) Micropipets (0.2 – 1 ml)

Mass or Weight We will always use the measurement of grams The scales we have are very accurate and expensive

Centrifuge Provide a centrifugal force to move liquid to the bottom of a tube Regular Micro

Aseptic Technique Microbes are ubiquitous Reduces the chances of contaminating yourself, others, or the experiment Involves the following: Sterilizing all equipment and supplies Keeping all equipment and supplies sterile Never touch anything with your bare hands Everything that touches an experimental substance should be sterile Keeping containers closed as much as possible Disinfecting your hands and bench frequently

Aseptic Technique What happens when you are not sterile? Lets see with the use of a “deadly virus” Everyone grab 1 tube of powder Glove up Squirt some glycerol on your glove and really rub it in Pour your tube of powder all over your hand Shake hands with 3 people Spread out and shake with 3 people not around you View your hand under UV light What happened?

Lesson 2: Essential Question How do we convert grams, meters, and liters correctly?

Lesson 2: Learning Objectives Know how to convert grams, meters, and liters correctly

Conversions

Conversions When you go to a smaller measurement, you move the decimal to the right When you go to a larger measurement, you move the decimal to the left Lets try these!

Lesson 3: Essential Question How do we properly use a glass pipet to transfer liquids?

Glass Pipets (1-25 ml) 4 Different Sizes Attach the Pipet and adjust volume by moving the wheel Watch closely while pipeting Must be disposed of in the sharps container

Lab Practice using graduated cylinders, scales, and glass pipets Follow the protocol given to you Make sure to be as accurate as possible and use aseptic technique at all times Dispose of everything in its proper place Don’t forget to clean up when you are finished!

Lesson 4: Essential Question How do we use a micropipet correctly?

Lesson 4: Micropipets

How to Use a Micropipet To draw up a solution Rotate the volume adjuster to the desired volume Place the appropriate tip on the barrel Press the plunger to the first stop Insert the tip into the solution Draw up the solution by gradually releasing the plunger Check to see that there are no air bubbles present in the tip

How to Use a Micropipet To expel the solution Touch the tip against the side of the tube or container into which the solution will be expelled Press the plunger to the second stop to make sure everything is expelled Remove the pipette tip from the container with the plunger still pressed Release the plunger slowly Eject tip into the appropriate waste container by pressing the tip-ejection button

Care of the Micropipet The following rules should be observed at all times: Never place a pipette near the edge of the bench where it can get knocked over Never rotate the adjuster below or beyond the designated ranges Never immerse the pipette barrel in liquid Never lay the pipette down or hold it horizontally when there is liquid in the tip Always wipe the pipette barrel when you are done Always use the appropriate tip for each pipette

Lab Practice using the micropipet Follow all instructions carefully Be sure to use proper aseptic technique at all times

Lesson 5: Essential Question How do we make solutions in the biotechnology laboratory?

Math Skills Needed for Biotech You must know how to calculate solutions using the following equations! You will do this every day in class Like conversions, they will not go away, YOU MUST LEARN THEM NOW!!!!! C1V1=C2V2 Mass/Volume

Solute/Solvent= Concentration Solutes (usually measured in g) Solvents (usually measured in ml) Mixed together until solute dissolves in solvent solute solution concentration =

C1V1=C2V2 (starting concentration)(starting volume)=(final concentration)(final volume) Solve for any of the variables, as long as you know the other 3 How do we actually make the solution once we have the answer?

Mass/Volume Equations The mass of solute in the amount of solvent Calculates the amount of solute needed per amount of solvent Mass/volume ___________ _____________ __________________ concentration desired X volume desired = mass of solute needed % Mass/volume __________ ____________ _________________ conc desired (decimal) X volume desired = mass of solute needed

Examples: Mass/Volume To make 100 mL of a 5 mg/mL hemoglobin solution ___________ _____________ _________________ concentration desired X volume desired = mass of solute to be weighed 5 mg/mL 100 mL 500 mg = 0.5 g concentration desired X volume desired = mass of solute to be weighed

Examples: % Mass/Volume To make 100 mL of a 3% CuSO4 solution 3 % = 0.03 __________ ____________ _________________ conc desired (decimal) X volume desired = mass of solute needed 0.03 g/mL 100 mL 3 g CuSO4 conc desired X volume desired = mass of solute needed