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DNA Fingerprinting TOOLS AND TECHNIQUES Intoducing the microLiter! A TINY amount…a millionth of a Liter Very difficult to measure because it is SOOO small Incredibly accurate when measured correctly Tools that measure these amounts are therefore INCREDIBLY expensive…be VERY careful with them!!! If a milliliter (mL) is 1/1000 of a Liter… A microliter (μl) is 1/1000 of a millileter (mL) The greek letter “mu” is used to represent micro (μ) Let’s PRACTICE Work through the second page of the packet “DNA Fingerprinting – Practice Worksheet” The TOOLS DNA Fingerprinting makes use of tools that can work with incredibly SMALL volumes We only need a very small sample of DNA We can run hundreds or thousands of samples at once in small spaces We can be INCREDIBLY accurate and precise We can use robotic assistance to run the full analysis The Micropipette Are like a normal pipette…but better They can hold incredibly small amounts of liquid with incredibly high accuracy and precision This makes them VERY expensive!!! Two types that we will use: P20: Can hold up to 20 μl P200: Can hold up to 200 μl Draw and label a picture to refer back to: Setting the Micropipette Each micropipette has a dial for its particular volume maximum NEVER set the dial above the MAXIMUM listed, this will damage the micropipette For a p20…NEVER set above 20 The three numbers represent the three place values Starting with the largest POSSIBLE place value Let’s PRACTICE with the second half of practice worksheet Techniques for Using Tips NEVER TILT THE PIPETTE!!! NEVER USE A NAKED PIPETTE!!! We use tips to hold liquid, not the pipette itself Liquids can actually damage (very severely) the internal parts To pick up a tip, push the pipette into the tip holder ALWAYS keep the tip box COVERED This keeps out contaminants After using ANY liquid, we will switch tips This will also help minimize the risk of contamination through mixing of samples To eject a tip, push down on the SIDE button This is attached to the metal rod that will push down on the tip Always eject tips into a WASTE BIN Techniques for using a Micropipette Push the PLUNGER to the FIRST Stop DO NOT GO FURTHER THAN THE FIRST STOP We must create a vacuum in order to pick up liquid While holding it down, insert the tip into your liquid DO NOT LET GO UNTIL YOU ARE IN LIQUID This will pick up air, not the liquid Release the plunger to pick up the liquid To eject, move the tip into the desired container Hold the tip against the inside surface of the container you wish to eject into Push the PLUNGER to the SECOND Stop to eject ALL liquid Let’s practice! Get a pipette each and practice picking up and ejecting a tip, and pushing the plunger to the FIRST and SECOND stop Using the microcentrifuge NEVER LOAD IT SIDEWAYS Like a washing/drying machine, it will spin out of control and potentially break ALWAYS balance the load In a 6-holder, this means you can load… NOT One Two Three Four NOT Five Six Balance the epitubes in the rotor Let’s practice!!! Do activity One and Two on the second instructional handout Answer the first two questions by converting Answer questions 3 and 4: the “Lab Observations” We will discuss and answer remaining questions after returning from break