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blood collection for control experiment was equally undertaken for the purpose with regards to the manufactures protocol . the sampling of the genomic DNA was used as templates for the PCR amplification with specific T .cruzi Mdna primers . Tcz1 and TC z2 and the KDNA primers s35 and s36 . the amplification reaction was run with 200ng templates DNA UNDER the subsequent conditions.0.2 Mm of each primer , 2.5 U Taq DNA Polymerase , 0.2 mM dNTP and 2 mM Mg Cl2 in a 25 ml final volume .
the triplicate amplification reaction were performed using the recommendations temperatures for Ndna ( 950C for five minutes , 30 cycles of 950 C for 30 s , 620C for 1 min nd 72 0C for 1 min and final extension at 72 o C for 5 min ) . The amplicons were resolved in the 1.3 % agarose gel The tumor tissue average RFU 0.254 corresponds to 406.25 standard and controls tissue average RFU 0.197corresponds to 275 standards. The curve is gently rises in a direct proportional manner to huge figures.
The scale of the graph presents 1 cm representing 125 ng/ul in y-axis while the 1 cm representing the 0.1 in x-axis (Maria & Frank , 10). 22. It is important to remove the supernatant then dry the pellet at the prevailing room temperature. Normally, the DNA do not attach or stick as expected on the walls of the tube following 70% ethanol wash. Caution and extreme should be maintained while to bid avoid any aspirating the pellet In conclusion, the protocol for DNA extraction and the PCR based typically on the main methods takes subsequent steps.
In the process of extraction large quantities of the DNA may be necessitate heating briefly at 65oC) for the suspension. The resulting large great molecular weight may undergo several days for proper
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