Worksheet # 1 Answers

PHR 647

 

Nucleic Acid Biotechnology Techniques 

1) Southern hybridization entails: 
a) digesting DNA molecules with restriction enzymes
b) separating the fragments gel electrophoresis
c) hybridizing the separating fragments to a radiolabelled DNA probe, 
d) and visualizing the specific fragment by autoradiography

2) The three steps in Polymerase Chain Reaction (PCR): 
a) Melt double-stranded DNA molecules by heating
b) Anneal short oligonucleotide primers by cooling
c) Synthesize a new DNA strand using Taq polymerase

The three components crucial for carrying out a PCR reaction: 1) DNA template 2) Primers 3) Taq polymerase 

3) DNA sequencing is a technique used for determining the order of bases in a DNA molecule. 

4) DNA libraries are a collection of DNA fragments. The two types of DNA libraries consist of: 
a) genomic libraries generated from digesting DNA molecules with restriction enzymes
b) cDNA libraried generated from mRNA molecules by copying the mRNA sequence into
a DNA sequence using reverse transcriptase

5) Indicate all techniques (a-d) that can be used to accomplish each application listed (I-IV):

a) Southern hybridization 
b) Sequencing 
c) Polymerase Chain Reaction 
d) Cloning 

I. Amplification of DNA sequences. c & d 
II. Diagnostic analysis of base mutations in DNA. a & b 
III. Production of a recombinant genetically engineered protein.
IV. Isolation of a new gene that was previously unknown. d