Transformation of Bacteria Lab

Jake Stewart
Period 1
May 27, 2016

Introduction
In my own words, transformation can be best described as a cell being genetically changed
due to of adding recombinant DNA (1). Plasmids are genetic structures in a cell that are
able to independently replicate of the chromosomes, normally in a tiny circular DNA strand
in the cytoplasm of a bacterium or a protozoan. In this lab, genes are being inserted into the
plasmids, and depending on whether the plasmids accept or reject them, they could
possibly transform. Recombinant DNA is DNA that has been created artificially by combining
elements from different organisms (2). I based my personal definitions of these words on
definitions that I found on Wikipedia. The purpose of this lab is to transform regular bacteria
into glow in the dark, ampicillin resistant bacteria. We are trying to see if the glow in the dark
gene and the ampicillin resistant gene will have any affect on the plasmids in the positive
and negative tubes. My hypothesis is that we will see growth in the Amp+, No Amp+, & No
Amp- petri dishes, after the altered plasmids are inserted into them.

Materials

Experimental

Control

-Two separate tubes that contain the following:

-Tube 1 (Experimental)
-bacteria
-calcium
-chlorite
-recombinant DNA (Plasmid DNA)
-Tube 2 (Control)
-bacteria
-calcium
-chloride
-Four petri dishes that are labeled and contain the following:
-Amp+
-Amp-No Amp+
-No Amp-Inoculating loops
-Pipettes
-Cup of ice water
-Small metal balls

Procedure
(See lab manual)

Results
The results of this lab were not what we expected, going into it. My hypothesis had been
that our group would see growth in the Amp+, No Amp+, & the No Amp- petri dishes and
that the Amp- dish would not grow at all. My hypothesis was wrong. Out of the four petri
dishes, only the Amp+ grew, and even though it did grow, it was only a subtle amount of
growth. The Amp-, the No Amp+, & the No Amp- petri dishes did not grow at all or change in
any way. Many others had similar hypothesis to mine, and many of them got similar results
to me.

Discussion
The results of our lab had been different than we had expected. Only the Amp+ petri dish
showed signs of growth. It was a bit hard to pinpoint the reason that our results had not
turned out the way we had expected them to. We had followed the procedure, and used the
right amounts of each substance. I believe the reason our results did not turn out they way
they were supposed to is because we did not hold the plasmid tubes in the ice water long
enough. The lab instructions said we were to hold the plasmid tubes in the water for fifteen
minutes, but we were short on time so we held them in for somewhere between four to five
minutes. That was eight to nine minutes less than we were supposed to hold them in for
which is a significant drop in time and that could possibly have been a crucial step to take if
we had wanted our results to turn out the way we had hypothesized.

References
(1)
(2)

https://en.wikipedia.org/wiki/Transformation_(genetics)
www.news-medical.net/life-sciences/What-isRecombinantDNA.aspx