Polymerase Chain Reaction PCR Reaction Mix PCR Reaction

  • Slides: 6
Download presentation
Polymerase Chain Reaction

Polymerase Chain Reaction

PCR Reaction Mix PCR Reaction §Template DNA §PCR Buffer §d. NTP (A, T, C,

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

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

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)

Polymerase Chain Reaction (PCR) 35 cycles 1 2 4 8 16 34, 359, 738,

Polymerase Chain Reaction (PCR) 35 cycles 1 2 4 8 16 34, 359, 738, 368