Biomolecular Techniques

Pre-Lab Questions
1. Transformation, conjugation, and transduction were discovered in the laboratory. How

important are these mechanisms of genetic recombination in nature?

2. Use the internet to investigate DNA fingerprinting. What is this process, and how is it

used in criminal forensics?

DNA fingerprinting is a technique used to establish a link between biological evidence and a

suspect. A DNA sample taken from a crime scene is compared with a DNA sample from a

suspect to make a conviction.

3. How can plasmids be used to manufacture proteins such as insulin for diabetic patients,

or antibiotics for bacterial infections?

4. Research a genetic disease that may be cured through the use of genetic engineering.

What techniques are being explored to correct the problem? How do concerns of

designer embryos stem from this technology?

Gene replacement therapy starts with scientists making a new copy of a gene that is not working or not
there. Such as autism, autism can be somewhat cured be using gene replacement therapy.

Experiment 1: DNA Extraction

Post-Lab Questions
1. Which DNA bases pair with each other?

Cytosine pairs with guanine and adenine pairs with thymine. 

 2. How is DNA information used to make proteins?

3. What is the purpose of the following reagents in the experiment:

a. Salt (in the DNA Extraction Solution):

By adding salt, we neutralize the DNA and make the molecule less hydrophilic, meaning it

becomes less soluble in water.

b. Detergent (in the DNA Extraction Solution):

It acts the same way in the DNA extraction protocol, pulling apart the fats of the body.

c. Ethanol:

The role of the ethanol is to remove the solvation shell surrounding the DNA.

4. What else might be in the ethanol/aqueous interface? How could you eliminate this?

When you add the solution with cold ethanol 70% you would have a mix of DNA, RNA and other

soluble salts.

5. What is the texture and consistency of the DNA?

The texture and consistency of the DNA is thick, cloudy, somewhat “stringy”. 

6. Is the DNA soluble in the aqueous solution or alcohol?

DNA is soluble in an aqueous solution.

Experiment 2: Restriction Enzymes

Post-Lab Questions
1. How can DNA be fragmented into very specific sections?

2. Where do restriction enzymes come from? What is their function in nature?

3. How do molecules of varying sizes separate in electrophoresis? What is the purpose of

the gel? What about the electricity?

4. Investigate one way in which electrophoresis is used in medicine today. Write

one or two paragraphs to explain your findings.

Experiment 3: Cloning a DNA Fragment into a Bacterially-Derived Plasmid
Vector
Data
Record your data in this section.

Post-Lab Questions
1. What is the expected size of the plasmid plus the cut foreign DNA?

2. What type of ends do the enzymes BamHI and EcoRI produce? How does this type of

end facilitate cloning?

3. What enzyme is necessary to permanently link the digested foreign and plasmid DNA

together to form the recombinant DNA molecule? How does this enzyme work?

4. How would you clone a gene into a plasmid if there were no common restriction sites

between the two DNA sequences?

 
Experiment 4: Gel Electrophoresis
Data Tables
Table 2: Electrophoresis of Dye Samples

Well Sample # of Bands Migration

1 Pyronin
2 Methyl Orange
3 Ponceau G
4 Xylene Cyanol
5 Unknown 1
6 Unknown 2

Post-Lab Questions
1. What is in each of the unknown samples?

2. Why is electrical current necessary to separate molecules using electrophoresis?

3. Why is agar an appropriate medium to use for separating molecules? Research another

type of gel and provide a brief explanation regarding why it could be used rather than an

agar gel?

4. What is the charge of the samples? What do you think would happen if the molecules

held the opposite charge?

5. How is electrophoresis similar to, and different from, chromatography?