Tran Nguyen
Professor Pate
BIO 180 Lab
CRN 60106
05/01/2018
Lab #10-12: DNA
Introduction
In lab #10, purified plasmid DNA (pAMP) is digested with some restriction enzyme
called Eco RI. Then, to separate and visualize the fragments of the DNA by size, gel
electrophoresis is used, and with the information obtained, a plasmid map is able to be
constructed.
“Bacteria transformation is defined as the uptake and expression of naked DNA by
bacterial cells.” In lab #11, the goal is to clone the Green Fluorescent Protein (GFP) – the glow
gene obtained from a jellyfish – on bacteria, which means making bacteria glow in the dark. For
this to happen, bacterial cells are transformed with pAMP/GFP, which also means digesting
pAMP/GFP using the techniques learned in lab #10. Finally, bacteria are plated and incubated at
37°C, and green colonies are grown in liquid culture.
Lastly, in lab #12, with the method of alkaline lysis (simplified version), plasmid DNA is
isolated, which means getting the DNA from the bacteria containing the plasmid pAMP/GFP
from lab #11.
1
, Tran Nguyen
Data and Results
Lab 10:
EcoRI
EcoRI/ Uncut EcoRI/ Bam/
Bam DNA Hind III Hind III
Figure 1. Electrophoresis Gel of Digested Plasmid with Restriction Enzyme
Lab 11:
Figure 2. Agar Plates of Transformed Bacteria Using UV Light
2
Professor Pate
BIO 180 Lab
CRN 60106
05/01/2018
Lab #10-12: DNA
Introduction
In lab #10, purified plasmid DNA (pAMP) is digested with some restriction enzyme
called Eco RI. Then, to separate and visualize the fragments of the DNA by size, gel
electrophoresis is used, and with the information obtained, a plasmid map is able to be
constructed.
“Bacteria transformation is defined as the uptake and expression of naked DNA by
bacterial cells.” In lab #11, the goal is to clone the Green Fluorescent Protein (GFP) – the glow
gene obtained from a jellyfish – on bacteria, which means making bacteria glow in the dark. For
this to happen, bacterial cells are transformed with pAMP/GFP, which also means digesting
pAMP/GFP using the techniques learned in lab #10. Finally, bacteria are plated and incubated at
37°C, and green colonies are grown in liquid culture.
Lastly, in lab #12, with the method of alkaline lysis (simplified version), plasmid DNA is
isolated, which means getting the DNA from the bacteria containing the plasmid pAMP/GFP
from lab #11.
1
, Tran Nguyen
Data and Results
Lab 10:
EcoRI
EcoRI/ Uncut EcoRI/ Bam/
Bam DNA Hind III Hind III
Figure 1. Electrophoresis Gel of Digested Plasmid with Restriction Enzyme
Lab 11:
Figure 2. Agar Plates of Transformed Bacteria Using UV Light
2
