Question 1: In experiment 4, what is in unknown 2? a: Methyl orange and pyronin

Question

Question 1: In experiment 4, what is in unknown 2?

a: Methyl orange and pyronin

b: xylene cyanol and pyronin

c: xylene cyanol and methyl orange

d: ponceau G and methyl orange

In experiment 4, what is in unknown 1?

methyl orange and ponceau G

methyl orange and pyronin

ponceau G and xylene cyanol

xylene cyanol and methyl orange

In experiment 4, which molecule is positively charged?

methyl orange

xylene cyanol

ponceau G

pyronin

a: Methyl orange and pyronin

b: xylene cyanol and pyronin

c: xylene cyanol and methyl orange

d: ponceau G and methyl orange

Lab 14 Biomolecular Techniques Experiment 4: Gel Electrophoresis In this experiment, you will prepare an agarose gel and use an electric current to separate dyes of different molecular weights and charges. You will also analyze two unknown dyes and determine their composition based on results from your known samples. You will then connect gel electrophoresis to forensics Materials (15 mL) 0.8% Agarose 2 Alligator Clips 9V Battery (1) 100 mL Beaker Carbon Fiber Fabric 100 mL Graduated Cylinder Methyl Orange 6 Micro-Tip Pipettes Mini Gel Electrophoresis Chamber with 6-Tooth Unknown #1 Unknown #2 Xylene Cyanol Pair of Disposable Gloves (located in your safety kit) Safety glasses (located in your lab safety kit) *Hot Pad or Towel *Microwave Oven or Boiling Water Bath Paper Towels Scissors Comb Ponceau G Pyronin Ruler (15 mL) 1X TBE Buffer T ransfer Pipete Water You Must Provide Procedure 1. Put on your safety glasses and a pair of disposable gloves (located in your lab safety kit) 2. Insert the gel chamber comb into the slots of one end of the gel box. The comb will cause the agarose to mold around its teeth and create wells once the gel has cooled and solidified. The teeth of the comb should touch the bottom of the chamber. Note: The end of the gel chamber that you insert the comb into will be referred to as the "top" end of the chamber and the opposite end will be referred to as the "bottom" end of the chamber (Figure 9) 264

Explanation / Answer

1. The order of the dyes is pyronin, methyl orange, ponceau G, xylene cyanol, unknown 1 and unknown 2. The unknown 2 can be mixture of dyes 1 and 4 (pyronin and xylene cyanol). Option b is correct.

2. The unknown 1 can be ponceau G and methyl orange which is a mixture of 2 and 3 dyes. Option a is correct.

3. The methyl orange is positively charged in acidic conditions but in alkaline condition the positive charge is neutralized. hence, unknown sample 2 is positively charged and migrates at negative end.

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