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The identification of the bacteria was performed using the standard method of comparing the sequence similarity of the 16S rDNA region [19].
Large-scale expression of protein in bacteria was performed using the procedures of Dr. H. S. Gill in The PEPCC Laboratory of Robotics at Case Western Reserve University.
In situ hybridization of bacteria was performed using the following oligonucleotides: EUB-338 5'-GCT-GCC-TCC-CGT-AGG-AGT-3' FITC-labeled (eubacterial probe; [79]) and Sau-69 5'-GAA-GCA-AGC-TTC-TCG-TCC-G-3' Cy3-labeled (S. aureus probe; [80]).
Fluorescent staining of bacteria was performed using a previously described method with slightly modifications.
Antibiotic susceptibility testing of isolated bacteria was performed using the traditional disc diffusion method.
Identification of bacteria was performed using conventional physiological and biochemical methods [ 13, 14].
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Production of most of the extracellular enzymes by microbes (fungi, yeast and bacteria) are performed using submerged fermentation technique, which provides various advantages including medium sterilization, cultivation of cultures due to easy monitoring and controlling of parameters like nutrients concentration, temperature, aeration, pH and moisture, and end product purification.
Antimicrobial susceptibility tests of CUR-I and antibiotics against five diarrhea causing bacteria were performed using the standard broth microdilution method.
To confirm the recovered bacteria, PCR was performed using the specific primers to S. pneumoniae (5'-TGTTGCCAGCTTGTTCTTAGGAGGA-3', 5'-TTTTGACGTCAACTCAGCTTCCGAT-3') [ 20].
To test the null hypothesis that essential oil of C. verbenacea had no effect on periodontal bacteria, intergroup analysis was performed using Fisher's exact test.
Transformation of WT Col-0 and co mutant plants with Agrobacterium bacteria carrying recombinant constructs was performed using the floral dip method [ 81, 82].
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