PCR Reaction Mix PCR Reaction §Template DNA §PCR Buffer §d. NTP (A, T, C, G) §Forward Primer §Reverse Primer §Polymerase Enzyme (Taq)
Polymerase Chain Reaction (PCR) 5’ – ACGTAGCGATGCTAGCTGACACTG – 3’ I I I I I I I Template DNA 3’ – TGCATCGCTACGATCGACTGTGAC – 5’ 5’ – ACGTAGCGATGCTAGCTGACACTG – 3’ Denature (96 o. C) Single PCR cycle 3’ – TGCATCGCTACGATCGACTGTGAC – 5’ 5’ – ACGTAGCGATGCTAGCTGACACTG – 3’ I I 3’ – TGTG – 5’ 5’ – GTACG – 3’ 3’ – TGCATCGCTACGATCGACTGTGAC – 5’ I I Primer Annealing I 5’ – ACGTAGCGATGCTAGCTGACACTG – 3’ I I I I I I I 3’ – TGCATCGCTACGATCGACTGTG – 5’ 5’ – GTACGTAGCGATGCTAGCTGACACTG – 3’ I I I I I I I I 3’ – TGCATCGCTACGATCGACTGTGAC – 5’ Extension(72 o. C)
Polymerase Chain Reaction (PCR) 5’ – ACGTAGCGATGCTAGCTGACACTG – 3’ I I I I I I I 3’ – TGCATCGCTACGATCGACTGTG – 5’ 5’ – GTACGTAGCGATGCTAGCTGACACTG – 3’ I I I I I I I I Template DNA I 3’ – TGCATCGCTACGATCGACTGTGAC – 5’ 5’ – GTACGTAGCGATGCTAGCTGACACTG – 3’ Denature (96 o. C) 2 nd PCR cycle 3’ – TGCATCGCTACGATCGACTGTGAC – 5’ 5’ – GTACGTAGCGATGCTAGCTGACACTG – 3’ I I Primer Annealing 3’ – TGTG – 5’ 5’ – GTACGTAGCGATGCTAGCTGACACTG – 3’ I I I I I I I 3’– CATGCATCGCTACGATCGACTGTG – 5’ Extension(72 o. C)