China
Africa
Explore chapters and articles related to this topic
Enrichment of Anaerobic Methanotrophs in Biotrickling Filters using Different Sulfur Compounds as Electron Acceptors
Published in Chiara Cassarini, Anaerobic Oxidation of Methane Coupled to the Reduction of Different Sulfur Compounds as Electron Acceptors in Bioreactors, 2019
The primer pairs used for bacteria were forward bac520F 5’-3’ AYT GGG YDT AAA GNG and reverse Bac802R 5’-3’ TAC NNG GGT ATC TAA TCC (Song et al., 2013). The following program was used: initial denaturation step at 94°C for 5 min, followed by denaturation at 94°C for 40 sec, annealing at 42 °C for 55 sec and elongation at 72°C for 40 sec (30 cycles). The final elongation step was extended to 10 min. 5 μl of the amplicons were visualized by standard agarose gel electrophoresis at the following conditions: 1% agarose gel, a running voltage of 120 V for 30 min, stained by gel red, and documented using a UV transilluminator fitted with a Gel Doc XR System (Bio-Rad, USA).
Bacterial contamination of neglected hospital surfaces and equipment in an Algerian hospital: an important source of potential infection
Published in International Journal of Environmental Health Research, 2022
Somia Saadi, Rachida Allem, Mohammed Sebaihia, Abdelaziz Merouane, Mohammed Bakkali
Full length of 16S rRNA gene (1550 bp) was amplified for all bacterial isolates using two universal bacterial primers 27 F (5ʹ-AGAGTTTGATCCTGGCTCAG-3ʹ) and 1492 R (5ʹ-TACGGYTACCTTGTTACGACTT-3ʹ) for PCR amplification (Weisburg et al. 1991). The 50 µL reaction mix included 1X Go Taq Buffer (Promega, USA), 2.5 mM MgCl2, 0.1 mM of each dNTP, 1 U hot start Taq DNA polymerase (Promega, USA), 0.5 µM each primer, and 5 µL of DNA template. The reactions were carried out in an Eppendorf Mastercycler gradient under following PCR thermal program: initial cycle for 10 min at 94°C, 35 cycles (denaturation: 45 s at 94°C, annealing: 1 min at 55°C, extension: 1 min at 72°C) and an additional cycle of 10 min at 72°C. Aliquots of 5 μL of each reaction were analyzed on 1% agarose gel with 1% of ethidium bromide (Gel Doc XR+ System, Biorad, USA), 100 bp ladder was used as DNA molecular weight standard in TAE buffer.
Investigation of two different size microplastic degradation ability of thermophilic bacteria using polyethylene polymers
Published in Environmental Technology, 2022
Ceyhun Akarsu, Sadin Özdemir, Yasin Ozay, Ömer Acer, Nadir Dizge
A fast DNA Spin Kit for Soil was used to isolate DNA from the samples (MP Bio, USA). A Qubit fluorometer (Invitrogen, USA) was used to measure the amount and purity of DNA extracted after DNA isolation. Using universal primers PA5'AGAGTTTGATCCTGGCTCAG-3'PH5'AGGGAGGTGATCCAGCCGCA-3’, PCR analysis was used to amplify selected gene regions for species identification. The 40 µL PCR mixture contained 1.5 mM MgCl2, 0,2 mM dNTP mix, 1 ɥM each primer, 1U i-StarTaqTM DNA polymerase (INTRON Biotechnology, Inc, USA), and 10 ng of DNA template. A C1000TM Thermal Cycler (BIORAD, USA) PCR equipment was used for PCR amplification, which was programmed as follows: initial denaturation at 95 oC for 5 min, followed by 35 cycles of denaturation at 95 oC for 30 s, annealing at 55 oC for 30 s and extension at 72 oC for 45 s, with a final extension performed at 72 oC for 10 min. After the polymerisation reaction, the PCR product was purified with a QIAquick PCR Purification Kit (Qiagen, Germany) and measured with an Invitrogen Qubit fluorometer (USA). Gel Doc (BIORAD, USA) imaging equipment was used to take photographs of the PCR products after electrophoresis on a 1.5% agarose gel with 1xTAE buffer at 100 volts (Figure 2). The 16S rRNA gene was sequenced by the Iontek Company using a PA primer and an AB1373 Automated Sequencer (Invitrogen, Carlsbad, USA) (Istanbul, Turkey).
High rates of Salmonella contamination in raw kibbe from commercial establishments: predominance of Salmonella Give
Published in International Journal of Environmental Health Research, 2021
Jacqueline Tanury Macruz Peresi, Ivete Aparecida Zago Castanheira De Almeida, Inara Siqueira De Carvalho Teixeira, Sonia Izaura De Lima E Silva, Rejane Alexandre Silva Graciano, Monique Ribeiro Tiba-Casas
The PFGE technique was performed according to the standard protocol of the PulseNet network (Hunter et al. 2005; CDC 2013b). Each agarose block containing total DNA from a bacterial isolate was placed in a tube containing a digestion solution with 30U of XbaI enzyme per isolate. The tube was then incubated in a water bath at 37°C for 18h. Separation of the DNA fragments was performed using 1% Seakem Gold agarose gel electrophoresis in 0.5X TBE. The S. enterica Braenderup H9812 strain was used as a molecular marker (Hunter et al. 2005). The electrophoretic run was performed in the CHEF-DR III system (Bio-Rad Laboratories) under the following conditions: initial time – pulse end 2.2–63.8 seconds, voltage – 6V, run time 18–19 hours. After the run, the gel was stained with ethidium bromide and the image was captured using the GEL DOC EZ system (Bio-Rad).