Biochemistry Lab Report BIOCHEMISTRY LAB REPORT 4 2023| PORTAGE LEARNING|WITH PROCEDURES, EXPLANATION AND FINAL RESULTS.HIGHLY VERIFIRD Experiment #: Lab 4 Title: Chromatography of molecules. Purpose: The goals for the student performing the experiment are 1. Explain the fundamentals of chromatography. 2. Demonstrate ion exchange and size - exclusion chromatography. 3. Separate a Real -World sample. The student should be able to use the techniques learned and be able to apply them to further use in the lab or in a chemistry class again. Students should be able to fully recall and demonstrate how to do these techniques when completed the lab. Procedure: Part 1: • Setting up the columns. o Notes: The pH of a solution can impact the stationary phase and the molecules to be separated. o 1. The chromatographic media is poured in as a slurry. o 2. The media is permitted to settle. o 3. Buffer (mobile phase) is permitted to flow through the column. • Procedure: o Step 1: Put on appropriate lab equipment based on what solutions and chemicals will be used in the lab that they are performing. Ex: Gloves, Lab coat, Closed toe shoes, Protective glasses, etc. Also have the appropriate materials needed for completion of the lab. o Step 2: Place glass wool in column so that the media has a barrier to keep it from eluting. o Step 3: Place column on stand. Be sure to label columns. o Step 4: With prepared media place in column and let it settle for a couple of minutes. o Step 5: Let some of the buffer for the columns run through so that they are in the gel. Never let the gel go dry, always keep the buffer above the gel. o Step 6: Your columns are not set up for separating test mixture compounds. Part 2: • Separation of two test mixtures. o Notes: ▪ There will be two test mixtures applied to each column. o Step 1: Put on appropriate lab equipment based on what solutions and chemicals will be used in the lab that they are performing. Ex: Gloves, Lab coat, Closed toe shoes, Protective glasses, etc. Also have the appropriate materials needed for completion of the lab. o Step 2: With columns already set up and have had buffer ran through Biochemistry Lab Report them know you are ready to begin separating test mixtures.
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