12. DNA Amplification with the Polymerase Chain Reaction (PCR) Make copies ( extend primers ) Starting DNA Template 5’ 5’ 3’ 3’ 5’ 5’ 3’ 3’ Add primers ( anneal ) 5’ 3’ 3’ 5’ Forward primer Reverse primer Separate strands ( denature ) 5’ 5’ 3’ 3’
13. In 35 cycles at 100% efficiency, 1.07 billion copies of targeted DNA region are created PCR Copies DNA through Multiple Thermal Cycles Original DNA target region Thermal cycle Thermal cycle Thermal cycle
19. Distribution of different genotypes of FOKI polymorphism & alleles frequency in patients & controls: FF genotype Ff genotype ff genotype Allele (%) F f Patients (n=50) N (%) 34 (68) 9 (18) 7 (14) 72.9 27.1 Controls (n=20) N (%) 20 (100) 0 (0) 0 (0) 100 0
20. Distribution of different genotypes of BSMI polymorphism & alleles frequency in patients & controls: BB genotype Bb genotype bb genotype Allele (%) B b Patients (n=50) N (%) 27 (54) 15 (30) 8 (16) 64.6 35.4 Controls (n=20) N (%) 1 (5) 2 (10) 17 (85) 13.6 86.4
21. Comparison of the studied parameters in FOKI 0.07 0.68±.088 .74 ± .140 BMD lumbar (g/cm 2 ) 0.000 0.68±.040 0.77 ± .129 BMD femoral (g/cm 2 ) 0.04 -2.11 ±0 .91 -1.54 ± 1.14 T score lumbar 0.3 -1.21 ± 1.14 -.94 ± .983 T scorefemoral P-value Ff+ff (N=16) Mean ± SD FF (N=54) Mean ± SD Variables
27. O.R ratio of FOK1genotypes of patients and controls: (32) (0) (%) 16 0 Count Ff+ff (68) (100) (%) 1.588(1.29-1.95) 0.004 34 20 Count FF cases control FOK1 OR (95% CI) P value GROUP
28. O.R ratio of BSM1 genotypes of patients and controls: (16) (85) (%) 8 17 Count bb (84) (15) (% ) 29.75(7.04-125.8) 0.000 42 3 Count BB+Bb cases control BSM1
extraction of DNA using salting out technique followed by amplification of extracted DNA at exon 2 for FOKI & intron 8 for BSMI and amplified DNA was then cut by restriction enzyme FOKI for detection of FOKI polymorphism and by enzyme BSMI for detection of BSMI polymorphism and then electrophoresis of the DNA was done on agarose gel.
![[object Object]](https://image.slidesharecdn.com/patternofvitamindreceptorpolymorphism-100625104053-phpapp02/85/Pattern-of-vitamin-d-receptor-polymorphism-1-320.jpg)
![Introduction ,[object Object]](data:image/gif;base64,R0lGODlhAQABAIAAAAAAAP///yH5BAEAAAAALAAAAAABAAEAAAIBRAA7)