Lexi S.P.

Ms. Mathews
STEM 1/2 period
23 March 2018
pGLo Lab Report
Purpose:
The purpose of this lab was to learn about genetically modified organisms(GMOs)
and how they work. To do this we inserted the pGlo gene into e.Coli to make it
glow under UV light.
Hypothesis:

Plate + DNA - DNA

LB Yes​(should grow) Yes
No​(should not glow) No
LB/amp Yes No
No No
LB/amp/ara Yes
Yes
Grow
glow
Procedure:
In this lab we used th Babec pGLo transformation instructions “pGLo
Transformation.” ​BABEC: Bay Area BIoscience Education COmmunity,
babec.org/gfp-transfcormation/.
Data:

1. LB/amp/ara plate 2. LB/amp plate with 3. LB/amp plate with

with +DNA -DNA +DNA

4. LB plate with +DNA 5. LB plate with -DNA

Analysis:
The plates of e.Coli above contain Either/or/all/some of the components
following; LB(helps grow bacteria), amp(ampicillin which kills the bacteria
without the pGlo gene because pGlo is immune to ampicillin, and ara(arabinose
which is a sugar that turns on the pGlo gene). The arabinose and ampicillin have to
work together to bring RNA Polymerase to start transcription and produce the
fluorescent green protein. We inserted the pGlo plasmid into the +DNA so it would
grow in every plate but the -DNA doesn’t have this plasmid so the amp will kill it.
Plate 1 above contains all three. The colonies shown have grown because of
the LB, and there aren’t as many colonies as plate 4 and 5 because of the
ampicillin. IT also has a faint glow because of the arabinose which activated the
glow. Plate 2 has LB and amp, there was no growth of the e.Coli as expected
because the amp killed it off, but there was a colony of growth on the side probably
due to contamination. Plate 3 also had LB and amp but because it had +DNA, with
the pGlo plasmid resistant to amp, it could grow and the amp. But since there was
no ara again it couldn’t glow. Plate number 4 and 5 only had LB so none of the
bacteria was killed off making there be a significantly higher number of colonies
that grew. It also didn’t glow there was no ara or amp to activate it, this was
expected.
This information proves our hypothesis to be correct for the most part. The
first plate should have glowed brighter if we could have waited a little longer for it
to grow. The second plate also shouldn’t have had a growth but this was most
likely do to contamination, not properly or carefully sterilizing the pipets. Other
than these two errors everything turned out as expected.
Some improvements could be to be more organized and prepared to prevent
contamination and the water not being at the right temperature, which would make
sure that our timing was precise. The lab also seemed to be a little rushed due to
the amount of plates we had to do in the time given.
This lab could lead to further investigating, such as learning how we can
make the e.Coli glow in different colors.
CLEAR paragraph:
We transformed the e.Coli to glow fluorescent green under UV light. We
inserted the pGlo plasmid into the e.Coli DNA.Then we made 5 different plates: 1
with LB, amp, and ara; 2 with LB and amp; and 2 with LB. After we went through
the proper procedure to to see the results our hypothesis proved to be true.We
know that you nee LB, amp, and the ara to produce fluorescent green bacteria
because the other plates never had ara, you need amp and ara to activate the pGlo
gene, and did not glow. This proves that we transformed the DNA, the former
e.Coli couldn’t glow but now that we inserted the pGlo gene it glows. We know
that our’s was glowing because when we put it under the UV light we saw a faint
green glow from the colonies of bacteria. We successfully changed the DNA of
e.Coli to glow.