Worksheet # 1

PHR 647

 

Nucleic Acid Biotechnology Techniques 

1) Southern hybridization entails: 
a) digesting _____________ molecules with ______________ _____________, 
b) separating the fragments by __________________, 
c) hybridizing the separating fragments to a __________________ probe, 
d) and visualizing the specific fragment by __________________. 

2) The three steps in Polymerase Chain Reaction (PCR): 
a) ______________ double-stranded DNA molecules by _______________. 
b) ______________ short oligonucleotide primers by _______________. 
c) ______________ a new DNA strand using _________ _______________. 

The three components crucial for carrying out a PCR reaction: 
1) ______________ 2) ______________ 3) ______________ 

3) DNA sequencing is a technique used for determining the ____________ of _________ in a DNA molecule. 

4) DNA libraries are a collection of DNA fragments. The two types of DNA libraries consist of: 
a) ____________ libraries generated from digesting DNA molecules with ______ _____________. 
b) ___________ libraried generated from mRNA molecules by copying the mRNA sequence into a DNA sequence using ______________
_____________. 

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. _______________ 
II. Diagnostic analysis of base mutations in DNA. __________ 
III. Production of a recombinant genetically engineered protein. ________ 
IV. Isolation of a new gene that was previously unknown. ____________