3.1.

1 DNA extraction This is the technique by which deoxyribonucleic acid is isolated from other cellular materials for purposes of analysis and use in biotechnological research (that is nucleic acid based diagnosis and cloning experiments). DNA was used for research, making a clonal gene bank, cloning into expression vectors for protein expression designing of immunological target epitopes for vaccine and or drug. 3.1.1.1 Principle DNA was extracted from the FTA cards, when the infected spotted blood in the FTA card is immersed in the chelex buffer, and then heated to 94C; the double stranded DNA in the card is denatured producing two single stranded DNA molecules which have less affinity for the card and thus they are displaced by the chelex reagent. The chelex and the FTA card are then removed and discarded and the DNA harvested for use. 3.1.1.2 Requirements Blood spotted FTA cards Chelex reagents Cutting tool Tris EDTA (TE) buffer Pipette tips PCR tubes PCR machine pippettes

3.1.1.3Procedure Three 1.2mm discs were punched out of the card using a cutting tool and put in a plastic tube. Tris EDTA buffer was added to the samples and incubated for five minutes at room temperature; TE is used to hydrate the DNA in solution. The samples were washed twice using TE buffer and stored at 40c until required for further analysis. 3.1.1.4 Results and interpretation To confirm if it was really the DNA of interest extracted, it was amplified using PCR technique and then run on Agarose gel alongside a known standard marker. 3.1.1.5 Quality assurance aspects Sterility was mandatory to prevent contamination of the extracted DNA with external or environmental DNA. It was essential to time the incubation time of the disc with chelex as if the discs take long in the buffer chelex can re-enter the buffer and displace DNA back into the discs.