Rainbow Dye Electrophoresis to Identify the Molecules in a Mixture - - PowerPoint PPT Presentation

Lab 4: Molecular Rainbow

Dye Electrophoresis to Identify the Molecules in a Mixture

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

Notebook

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?

Observations at 5 Minutes

• Write 2-3 sentences describing your
• bservations 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

Notebook