Introduction:

I am investigating the effect of heat shock on E. Coli colonies. If heat shock is induced for a
shorter period of time, then more E. Coli colony growth will be visible on the LB plate with
Amp. I predict that a longer heat shock will result in inhibited bacterial growth. This is because
heat induces alterations of E. Coli cells, like their membrane, cytoplasm, ribosome and more.
Experimental design:
The variable assigned to our group was the 90 second heat shock. For group 8, a few colonies
were collected from the E. Coli agar plate and resuspend in the 250ul calcium chloride solution.
This was kept in an ice bath for 5 minutes. Next, 10ul of DNA plasmid was added into the cell
suspension. This was mixed and set in an ice bath for 5 minutes. Then, the transformation mix
was heat shocked by adding it into a 45 degrees Celsius water bath for 90 seconds. It was then
returned to ICE for two minutes. In the next step, LB broth was pipetted into the transformation
mix, mixed, and then incubated for 30 minutes. Following this, the transformation mixture was
pipetted onto four different agar plates. Each plate was pipetted with 100ul of the mixture, and
the LB plate without antibiotics served as a control group.
Experimental Results:
Group 8: Colony Growth Group 3: Colony Growth (paired
group)

LB plate 0 0
LB with Amp 14 (90 second heat shock) 9 colonies (180 second heat
shock)

Table 1: This table demonstrates the differences in number of colonies between Group 8 and
Group 3.
Transformation Efficiency:
# 𝑡𝑟𝑎𝑛𝑠𝑓𝑜𝑟𝑚𝑎𝑛𝑡𝑠 (𝑐𝑜𝑙𝑜𝑛𝑖𝑒𝑠)∗𝑓𝑖𝑛𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒 (0.5𝑚𝐿) (14)∗(0.5)
Group 8: = = 700 𝐶𝐹𝑈/𝑢𝑔
𝑢𝑔 𝑜𝑓 𝑝𝑙𝑎𝑠𝑚𝑖𝑑 (0.1)∗𝑣𝑜𝑙𝑢𝑚𝑒 𝑝𝑙𝑎𝑡𝑒𝑑 (0.08𝑚𝐿) (0.1)(0.10)

# 𝑡𝑟𝑎𝑛𝑠𝑓𝑜𝑟𝑚𝑎𝑛𝑡𝑠 (𝑐𝑜𝑙𝑜𝑛𝑖𝑒𝑠)∗𝑓𝑖𝑛𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒 (0.5𝑚𝐿) (9)∗(0.5)

Group 3: = = 450 𝐶𝐹𝑈/𝑢𝑔
𝑢𝑔 𝑜𝑓 𝑝𝑙𝑎𝑠𝑚𝑖𝑑 (0.1)∗𝑣𝑜𝑙𝑢𝑚𝑒 𝑝𝑙𝑎𝑡𝑒𝑑 (0.08𝑚𝐿) (0.1)(0.10)

Conclusion:
The variable of 90 second heat shock versus 180 second had a significant effect on
transformation. As seen in Table 1 group 8, with a 90 second heat shock, experienced a higher
colony growth. In group 3’s LB with Amp plate, the colony growth was smaller as it experienced
a longer heat shock (180 seconds). These results support my prediction. My predictions state that
as the colonies experience a longer period of heat shock, there are less colonies that are on the
LB with Amp agar plate. As seen in Table 1, group 3 experienced a lower number of colonies as
they were exposed to the heat shock twice as long as group 8. In the calculated transformation
efficiency values, group 8 has a higher value which is proven by the experimental data collected.