ELN Lab 4 5-16.docx

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Microbiology and Cell Science
MCB 3020L

Chloe Evetts (8990-4674) Cameron Jacobs May 15, 2013 Pipetting Dilutions Purpose: The purpose of this experiment is to learn about pipetting and experimental design. By diluting a sample, we will learn proper pipetting technique, as well as how to properly dilute a sample. Hypothesis: By being able to properly use a pipette, we believe that a multitude of experiments can be performed, including the dilution of a sample. Procedure: 1. Take 1 8-well ELISA test strip (per student) 2. Label your strip on the side you start with your seat number (on the tab) 3. Use the P200 pipette, attach a white tip, fill well 1-8 with 90 μl H2O 4. Use the P20 pipette, attach a white tip and transfer 10 μl crystal violet (from your Gram stain kit) into well 1 and 10 μl crystal violet into well 2. 5. To perform your 1:100 dilutions, use a P20 pipette attach a white tip and transfer10 μl from well 1 into well 3 - mix, repeat and transfer 10 μl from well 2 to well 4 – mix after each transfer 6. To perform your 1:1000 dilution Pipette 10 μl from well 3 and add it to well 5 – mix, repeat and transfer 10 μl from well 4 to well 6 – mix after each transfer 7. Leave well 7 and 8 with water only (negative control) Give your strip to the TA and get your optical density (OD 600 reading. We will determine the OD with a spectrophotometer. Your TA may ask you to graph the data from your class to determine the quality of your pipetting and graphing skills. Results: Variable: concentration Positive control: pure crystal violet Negative control: pure wa
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