Used: i) polymerase chain reaction-restriction fragment length Large-scale samples are required toĪnalyze and reveal the relationships between disease and SNPs.Īt present, the following methods are typically A veryĪccurate prediction of diseases present may be obtained by (DNA) sequences of the genes, in particular with the SNPs.
These diseases are often associated with the deoxyribonucleic acid In children which is associated with IL-13 ( 1, 2) andĭisease ( 4, 5), human mental diseases ( 6, 7),Ĭhronic obstructive pulmonary disease ( 8, 9) and Incidence and prognosis of various major diseases, including asthma Polymorphisms have substantial clinical effects on the prevalence, Nucleotide polymorphisms (SNP) has been associated with The presence of specific human gene single The optimized PCR-SSP is suitable for polymorphism analysis of polygenic SNPs in large genomic DNA samples and a number of different genes. The cycling system comprised 5 start cycles and took 15 min to melt a genomic DNA sample using a touchdown protocol. The PCR-SSP amplification system was optimized in a 20 µl reaction system, the quantities of Mg2+, dNTPs, pfu Taq, primers, control primers and genomic DNA were 3.25 µM, 0.5 mM, 2.5 units, 0.5 µM, 0.2 µM and 0.15 µg, respectively. The optimized PCR-SSP method was used to analyze the polymorphisms of the following genes: mutations -308A/G and -238G/A in TNF-α, -174G/C in IL-6 and C/T mutation at exon 188 of CYP2D6 *10B. The resulting optimized reaction system was used to determine the melting temperature of the genomic DNA and the cycling parameters. The amplification system parameters, including the concentrations of Mg2+, dNTPs, pfu Taq, primers and control primers, were optimized using the designed PCR-SSP reactions. The present study aimed to explore the factors that affect polymerase chain reaction using sequence-specific primers (PCR-SSP) and to establish an optimized PCR-SSP method for detecting multiple gene polymorphisms simultaneously.
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