RAPD-PCR was performed using the extracted DNA to investigate the biotype diversity of P. multocida in the study isolates [33 (link),34 ]. The RAPD-PCR was performed using (5´-GCGATCCCCA-3´) primer following the previously optimized protocol for Escherichia coli [22 ]. Using previously published pathogenic gene-specific primers (Table-1 ) for PCR assays, we surveyed ten pathogenic genes in the strains of P. multocida [9 (link),16 (link)]. Briefly, the PCR mixer possessed 2 mL DNA template (300 ng/mL), 10 mL PCR master mix 2X (GoTaq® Colorless Master Mix, Promega, Madison, WI 53711 USA) and 1 mL (100 pmol/mL) of each primer (Table-1 ) in each reaction tube. The cycling condition for PCR amplifications was as follows: 94°C for 5 min; 35 cycles of 1min at 94°C, 1 min at 50-60°C, and 1 min at 72°C; and 72°C for 7 min. Amplified PCR products were visualized on 1.5% agarose gel prepared in 1× TAE buffer. After gel electrophoresis, the images were captured using Image ChemiDoc™ Imaging System (Bio-Rad, USA) [9 (link)].
Image chemidoc imaging system
Manufactured by Bio-Rad
Sourced in United States
The Image ChemiDoc™ Imaging System is a laboratory instrument designed for capturing and analyzing images of various biological samples, such as gels, blots, and plates. The system utilizes a high-resolution camera and specialized optics to capture images, and includes software for image processing and analysis.
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Lab products found in correlation
3 protocols using image chemidoc imaging system
1
Biotyping Pasteurella multocida via RAPD-PCR
2
Detecting Virulence Genes in APEC Strains
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We surveyed 13 virulence genes (VGs) that are usually studied in APEC strains. The selected genes included: eae (Intimin) [29 (link)], stx1 (shiga toxins 1) [29 (link)], stx2 (shiga toxins 2) [29 (link)], fimH (type 1 fimbriae) [32 (link)], hlyA (hemolysin) [32 (link)], papC (P fimbriae) [32 (link)], lt (heat-labile enterotoxin) [56 (link)], bfpA (bundle-forming pilus) [29 (link)], crl (curli fimbriae) [23 (link),31 (link),45 (link)], uidA (β-d-glucuronidase) [29 (link),32 (link)] aggR (aggregative adherence regulator) [57 (link)], ial (invasion-associated locus) [29 (link)], and cjrC (putative siderophore receptor) [29 (link),32 (link)]. Each PCR reaction contained 2 μL DNA template (300 ng/μL), 10 μL PCR master mix 2X (Go Taq Colorless Master Mix), and 1 μL (100 pmol/μL) of each primer in each tube. The PCR amplifications were conducted in thermocycler, and the cycling conditions were identical for all the samples as follows: Temperature at 94 °C for 5 min; 35 cycles of 1 min at 94 °C, 1 min at 50–60 °C, and 1 min at 72 °C; and 72 °C for 7 min. PCR amplicons were visualized on 1.5% agarose gel prepared in 1× TAE buffer. After gel electrophoresis, the images were captured using Image ChemiDoc™ Imaging System (Bio-Rad, Hercules, CA, USA) [29 (link),32 (link)].
3
Biotyping of Pasteurella multocida Isolates
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Random amplification of polymorphic DNA (RAPD)-PCR was performed with the extracted DNA to investigate the biotype diversity among the isolates [33, 34] . The RAPD-PCR was performed by using (5´-GCGATCCCCA-3´) primer following the previously optimized protocol for E. coli [31] . Using previously published pathogenic gene-specific primers (Table 1)
for PCR assays, we surveyed 10 pathogenic genes in the isolated strains of P. multocida [7, 19] .
Briefly, each PCR reaction contained 2 ߤL DNA template (300 ng/ߤL), 10 ߤL PCR master mix 2X (Go Taq Colorless Master Mix) and 1ߤL (100 pmol/ߤL) of each primer (Table 1) in each tube. The PCR amplifications were conducted in thermocycler, and the cycling conditions were identical for all the samples as follows: 94 °C for 5 min; 35 cycles of 1min at 94 °C, 1 min at 50-60 °C, and 1 min at 72 °C; and 72 °C for 7 min. PCR amplicons were visualized on 1.5% agarose gel prepared in 1× TAE buffer. After gel electrophoresis, the images were captured using Image ChemiDoc™ Imaging System (Bio-Rad, USA) [7] .
for PCR assays, we surveyed 10 pathogenic genes in the isolated strains of P. multocida [7, 19] .
Briefly, each PCR reaction contained 2 ߤL DNA template (300 ng/ߤL), 10 ߤL PCR master mix 2X (Go Taq Colorless Master Mix) and 1ߤL (100 pmol/ߤL) of each primer (Table 1) in each tube. The PCR amplifications were conducted in thermocycler, and the cycling conditions were identical for all the samples as follows: 94 °C for 5 min; 35 cycles of 1min at 94 °C, 1 min at 50-60 °C, and 1 min at 72 °C; and 72 °C for 7 min. PCR amplicons were visualized on 1.5% agarose gel prepared in 1× TAE buffer. After gel electrophoresis, the images were captured using Image ChemiDoc™ Imaging System (Bio-Rad, USA) [7] .
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