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Results:
Table 3: OD absorbance of
Spectrophotometer
Sample
OD
Fraction
Reading: 0
Mins
Filtrate
0.519
S1
0.839
Concentrat
ion
(mM)
1.0086 * 10-4
Velocity (
moles/min
)
3.3619 * 10-
0.994
-2.603*10-6
P1
1.767
1.685
-7.8091 * 10
S2
0.776
0.585
-1.81905*10- -6.063*10-6
P2
Malonate
0.533
0.803
0.544
0.524
1.04762*10-6 3.4921*10-7
-2.65714*10- 5
2.65714*105
Succinate
0.463
0.425
-3.61905*10
6
-1.206*10-6
Azide
0.670
0.731
5.80952*10-6 1.9365*10-6
Table 3: As discussed in methods, various concentrations of
Mannitol Assay Buffer, Azide, DCIP, Malonate, Succinate, and
subcellular fractions were added to the cuvettes (Table 1). The
cuvettes were then placed in a spectrophotometer and the OD
was read at 0 minutes of adding the sample fractions and at 3
minutes.
P2B:
The sample of P2, in the spectrophotometer with wavelength 600, had
the absorption of 0.647 at 0 mins and a 0.730 reading at 3 mins. The
DCIP reduction calculated for P2B was 2.6349 * 10-6 moles/min.
P2C:
The filtrate solution had an OD600 reading of 0.649 at 0 mins and a
0.676 reading at the 3 minutes mark. The reduction velocity of DCIP
was found to be 8.5714 * 10-7 moles/min.
S2 (Table 2):
The supernatant 2, in the spectrophotometer, had an absorption of
0.749 at 0 mins and a 0.948 reading at 3 mins, at 600nm wavelength.
The DCIP reduction calculated for S2 was 6.3175 * 10-6 moles/min.
Discussion:
Under the microscope, purple staining was observed for the F1
Solution. This Azure C purple staining is indicative of DNA presence
meaning that pieces of nucleus were present. The OD600 values for F1 at
zero minutes were 0.519 and 1.578 at 3 minutes mark. This high OD600
shows that mitochondrial succinate dehydrogenase was present.
S1 with the Azure C dye had a purple staining under the microscope.
This means that the supernatant 1 had traces of nucleus matter. The
OD600 at 0 mins was 0.839 and at 3 mins 0.994. The concentration was
less than F1, meaning that the concentration of succinate
dehydrogenase in this sample was low. This result suggests that
mitochondrial traces were present.
P1 had very few scattered pieces of purple staining in the solution. This
staining was observed under the microscope and suggests that there
was a very low concentration of DNA i.e. nucleus present. The OD600 at
0 mins was 1.767 and 1.685 at 3 mins. This observation suggests that
there was a very high concentration of succinate dehydrogenase. Most
of the P1 solution showed evidence of high mitochondrial
concentrations.
S2 had a 0.776 concentration at 0 mins and 0.585 OD600 at 3 mins.
These values suggest very low traces of mitochondrial substances. The
microscope created a green picture with what appears to be a large
stained area. This might suggest that nucleus material was present.
P2 had an OD600 reading of 0.533 at 0 mins and 0.544 at 3 mins. This
observation suggests that there was a very low if any concentration of
succinate dehydrogenase i.e. mitochondrial material.