Lab 4: Molecular Rainbow Dye Electrophoresis to Identify the Molecules in a Mixture
Notebook Lab Objectives • To understand that molecules vary in size, shape and charge • To analyze the results of molecule separation by gel electrophoresis • To assign relative size of dye molecules as a function of movement through the agarose gel • To determine the components of an unknown dye mixture • To predict how additional molecules would have migrated in gel electrophoresis based upon size and charge
Notebook Gel Electrophoresis Load and Run • Using a p20 micropipette, load 10 µL of each sample into the wells of an 0.8% agarose gel in the following order: Lane 1 – Bromophenol Blue (BB) Lane 2 – Methyl Orange (MO) Lane 3 – Ponceau G (PG) Lane 4 – Xylene Cyanol (XC) Lane 5 – Pyronin Y (PY) Lane 6 – Unknown 1 Lane 7 – Unknown 2 Lane 8 – Unknown 3 • Run at 125 volts for approximately 30 minutes
What do we know about gel electrophoresis? • Which molecule will migrate down the gel the farthest? 10,000 bp DNA piece 250 bp DNA piece 700 bp DNA piece
What do we know about gel electrophoresis? • Which molecule will migrate down the gel the fastest?
Notebook Observations at 5 Minutes • Write 2-3 sentences describing your observations at 5 minutes
Notebook Results
Notebook Results
Notebook Analysis • Respond to analysis questions #1-10 in your lab notebook • Write in full sentences and answer each question completely
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