Kibera is the largest informal settlement in Nairobi, Kenya and one of the largest in Africa and the world. It is located 7 km southwest of Nairobi and is home to around a million people and several animal species. Living conditions in Kibera are extremely poor and heavily polluted by human and animal waste. Say no to plagiarism. Get a tailor-made essay on “Why Violent Video Games Should Not Be Banned”? Get the original test Stool specimens were collected in sterile cups and kept at 4°C until tested. All study stool samples were stored at -80°C upon receipt at KEMRI laboratories in Nairobi. Extraction of Nucleic Material 180 to 220 mg or 200 μl of liquid stool samples were extracted using a modified procedure of the QIAamp Fast Stool Mini Kit where they underwent a lysate preparation process that included a step of mechanical disruption (bead beating), removal of inhibitors and elution of nucleic material using spin columns. The lysate was transferred to the spin columns and centrifuged. The columns were washed twice following the manufacturer's instructions; centrifugation at 14,000 RT-PCRSamples were tested for HEV by RT-PCR. Briefly, 25 µl reaction mixtures were prepared containing: 12.5 µL of 2x Taq man One-Step RT-PCR Master Mix Buffer, 1µl of 40x Multi Scribe and RNase Inhibitor Mix, 1µM forward primer, 1µM of reverse primer, 5.75µl of molecular grade water and 5µL of purified RNA. (Reactions were incubated at 50°C for 30 min, then at 94°C for 3 min. Thermal cycling was performed for 45 cycles at 94°C for 30 s, 42°C for 30 s, and 72°C for 30 s. a model 7500F thermal cycler (Applied Biosystems) Cycle threshold CT values ​​≤ 30 were taken into account for cDNA synthesis and amplificationA standardized internal cDNA synthesis and amplification protocol. one step was used to obtain amplicons for sequencing as follows: Keep in mind: This is just a sample. Get a custom article now from our expert editors. Get a custom essay A master mix containing 2 .0 μL of RNase-free water; 16 μL of 2X reaction mixture; 1 μL of 20 pmol/μL of forward and reverse primers; 1 μL of SuperScript III RT/Platinum Taq mix and 4 μL of the RNA template. The reaction mixture was incubated at 50°C for 45 minutes, then thermocycling was carried out for 40 cycles of 94 minutes. °C for 30 s; 42°C for 1 min and 60°C for 2 min in a Model 9700 thermal cycler (Applied Biosystems, Foster City, CA) (Oberste et al., 2006). Thermocycling was followed by a final extension at 60°C for 10 min. Reaction products were analyzed by electrophoresis in a 1% agarose gel and staining with 0.5 mg/ml ethidium bromide...