3.9 Methodology of Bacterial Identification
53
Agarose Gel Pic (1% agarose): Lane1 to Lane2: 1 kb ladder (1 µL), A1 (1 µL).
PCR Cycle:
95.0 °C—10 min
95.0 °C—30 s
54.0 °C—1 min 30 cycles
72.0 °C—1:30 min
72.0 °C—10 min
4.0 °C—
The results of phylogenetic tree are shown in Figs. 3.6 and 3.7.
3.10 Analytical Methods
3.10.1 Chemical Oxygen Demand (COD)
COD of the samples was measured following the principle of “Closed Reflux
Method.” At first, 2.5 mL of sample or its suitable aliquot was taken to a COD culture
tube and added with 1.5 mL of 0.1 N K 2 Cr 2 O 7 and 3.5 mL of Ag 2 SO 4 containing
H 2 SO 4 one after another. This culture tube solution was denoted as sample solution.
A similar tube was prepared taking 2.5 mL of distilled water in place of sample,
which is named as blank solution. Now, these two COD culture tubes were allowed
to digestion in a refluxing apparatus (COD Thermoreactor) at about (148 ± 2)º C for
2 h. After the digestion was over, the tubes were cooled and their contents are titrated
with 0.01 N ferrous ammonium sulfate (FAS) using ferroin indicator until a reddish
brown color appears. To determine the strength of FAS solution, a standardization
was also performed using a ‘blank’ tube as prepared earlier. In this case, the culture
tube was directly titrated with FAS without any digestion in the refluxing apparatus.
COD = {(A − B) × M × 8000/{volume of sample}
A = Volume of FAS for blank solution.
B = Volume of FAS for sample solution.
M = Calculated morality of FAS.
53
Agarose Gel Pic (1% agarose): Lane1 to Lane2: 1 kb ladder (1 µL), A1 (1 µL).
PCR Cycle:
95.0 °C—10 min
95.0 °C—30 s
54.0 °C—1 min 30 cycles
72.0 °C—1:30 min
72.0 °C—10 min
4.0 °C—
The results of phylogenetic tree are shown in Figs. 3.6 and 3.7.
3.10 Analytical Methods
3.10.1 Chemical Oxygen Demand (COD)
COD of the samples was measured following the principle of “Closed Reflux
Method.” At first, 2.5 mL of sample or its suitable aliquot was taken to a COD culture
tube and added with 1.5 mL of 0.1 N K 2 Cr 2 O 7 and 3.5 mL of Ag 2 SO 4 containing
H 2 SO 4 one after another. This culture tube solution was denoted as sample solution.
A similar tube was prepared taking 2.5 mL of distilled water in place of sample,
which is named as blank solution. Now, these two COD culture tubes were allowed
to digestion in a refluxing apparatus (COD Thermoreactor) at about (148 ± 2)º C for
2 h. After the digestion was over, the tubes were cooled and their contents are titrated
with 0.01 N ferrous ammonium sulfate (FAS) using ferroin indicator until a reddish
brown color appears. To determine the strength of FAS solution, a standardization
was also performed using a ‘blank’ tube as prepared earlier. In this case, the culture
tube was directly titrated with FAS without any digestion in the refluxing apparatus.
COD = {(A − B) × M × 8000/{volume of sample}
A = Volume of FAS for blank solution.
B = Volume of FAS for sample solution.
M = Calculated morality of FAS.