DNA sequencing was performed for confirmative detection of S. aureus strain; based on positive detection of femA and mecA, two isolates were obtained from humans and one from cattle, and the respective PCR products were purified from agarose gel. The purified femA and mecA fractions were sent to Bioneer, Korea, for DNA sequencing using Applied Biosystems DNA sequencing system. The genomic sequences were assembled and submitted in GenBank-National Center for Biotechnology Information followed by multiple sequence alignment using Basic Local Alignment Search Tool for phylogenetic tree construction and phylogenetic analysis by MEGA10 program (https://www.megasoftware.net/ ).
Dna sequencing system
Manufactured by Thermo Fisher Scientific
The DNA sequencing system is a laboratory instrument designed to determine the sequence of nucleotides in a DNA molecule. It provides the essential functionality to analyze the genetic composition of biological samples.
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Lab products found in correlation
2 protocols using dna sequencing system
1
Confirmative Detection of Staphylococcus aureus
2
Extracting Nematode DNA from FFPE Horse Tissue
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Genomic DNA was isolated from formalin-fixed paraffin-embedded (FFPE) tissue curls of the spinal cord from a horse. Eight 5 µm thick sections were cut from
FFPE tissues, placed in 1.5 ml tubes, deparaffinized with xylene, and washed with ethanol. Genomic DNA was extracted following the manufacturer’s instructions
(QIAamp® DNA FFPE Tissue Kit, Qiagen, Germany). PCR, targeting the 12S rDNA conserved among the nematode species Onchocerca
volvulus, Ascaris suum, and Caenorhabditis elegans, was performed with the DNA template using a primer set (F:
5′-GTT CCA GAA TAA TCG GCT A-3′, R: 5′-ATT GAC GGA TG (AG) TTT GTA CC-3;) designed by Casiraghi et al. [1 (link)]. PCR was performed at a final volume of 50 µl under the following conditions: 2 × EmeraldAmp® PCR Master Mix, 0.2 µM of each primer,
50–100 ng genomic DNA, and H2O up to 50 µl (EmeraldAmp® PCR Master Mix, Kusatsu, Japan). The following thermal profile was used: 40 cycles
of 94°C for 45 sec, 50°C for 45 sec, and 72°C for 90 sec. The 450 bp amplicon was submitted to Macrogen (Daejeon, Korea), where DNA sequencing was conducted
using an Applied Biosystems DNA sequencing system.
FFPE tissues, placed in 1.5 ml tubes, deparaffinized with xylene, and washed with ethanol. Genomic DNA was extracted following the manufacturer’s instructions
(QIAamp® DNA FFPE Tissue Kit, Qiagen, Germany). PCR, targeting the 12S rDNA conserved among the nematode species Onchocerca
volvulus, Ascaris suum, and Caenorhabditis elegans, was performed with the DNA template using a primer set (F:
5′-GTT CCA GAA TAA TCG GCT A-3′, R: 5′-ATT GAC GGA TG (AG) TTT GTA CC-3;) designed by Casiraghi et al. [1 (link)]. PCR was performed at a final volume of 50 µl under the following conditions: 2 × EmeraldAmp® PCR Master Mix, 0.2 µM of each primer,
50–100 ng genomic DNA, and H2O up to 50 µl (EmeraldAmp® PCR Master Mix, Kusatsu, Japan). The following thermal profile was used: 40 cycles
of 94°C for 45 sec, 50°C for 45 sec, and 72°C for 90 sec. The 450 bp amplicon was submitted to Macrogen (Daejeon, Korea), where DNA sequencing was conducted
using an Applied Biosystems DNA sequencing system.
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