Kibera is the largest informal settlement in Nairobi, Kenya, and one of the largest in Africa and the world. It is located 7 km south-west of Nairobi and is home to around one million people and several animal species. Conditions in Kibera are extremely poor and heavily polluted by human and animal rubbish and waste. Say no to plagiarism. Get a tailor-made essay on "Why Violent Video Games Shouldn't Be Banned"? Get an original assay Stool samples 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, Nairobi. Extraction of nucleic material 180 – 220 mg or 200 µl of liquid stool samples were extracted using a modified QIAamp Fast Stool Mini Kit procedure in which they were subjected to a lysate preparation process that included a mechanical disruption step (beating of the spheres), removal of inhibitors and elution of nucleic material using spin columns. The lysate was transferred to spin columns and centrifuged. The columns were washed twice following the manufacturer's instructions; centrifuged at 14,000Xg for 1 minute and eluted with 100 µl total nucleic acid (TNA) (Liu et al., 2013). RT-PCRS samples were tested for HEV by RT-PCR. Briefly, 25 µl of reaction mixes were prepared containing: 12.5 µl of 2x Taq man one-step RT-PCR master mix buffer, 1 µl of 40x Multi Scribe and RNase inhibitor mix, 1 µM of forward primer , 1 µM reverse primer, 5.75 µl molecular grade water and 5 µl purified RNA. (Reactions were incubated at 50°C for 30 min followed by 94°C for 3 min. Thermocycling was performed for 45 cycles at 94°C for 30 s, 42°C for 30 s, and 72°C for 30 s cycle threshold CT values ​​of ≤30 were taken into account in a model 7500F thermal cycler (Applied Biosystems). and one-step cDNA amplification: Please note: This is just an example Get a custom paper from our expert writers now Get a custom essay A master mix containing 2.0 μL of RNase-free water 16 μL of reaction mix. 2X 1 μL of 20 pmoles/μL Forward and Reverse Primers (292 and 222 Table 1); 1 μL of SuperScript III RT/Platinum Taq Mix and 4 μL of RNA Template The reaction mixture was incubated at 50°C for 45 minutes at 94°C for 3 minutes, then thermocycling was performed for 40 cycles at 94°C for 30 s; 42°C for 1 minute and 60°C for 2 minutes 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 minutes. The reaction products were analyzed by 1% agarose gel electrophoresis and staining with 0.5 mg/ml ethidium bromide..