Application of orthogonal design to optimize real-time quantitative PCR conditions
- VernacularTitle:正交设计在实时定量PCR检测条件优化中的应用
- Author:
Qiuxiang DU
;
Junhong SUN
;
Xiaowei WANG
- Publication Type:Journal Article
- Keywords:
Forensic pathology;
Orthogonal design;
Real-time Quantitative Polymerase Chain Reaction;
Skin injury;
ICAM-1
- From:
Chinese Journal of Forensic Medicine
1986;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the application of orthogonal design to optimize real-time quantitative PCR conditions and perform it for ICAM-1.Methods After model of a skin injury was established in rats,total RNA was isolated from the injured skin tissue and reverse transcriptase reactions were carried out.Using orthogonal design real-time quantitative PCR conditions,the expression level of ICAM-1 gene was studied by use of cDNA as template.The standard of score was set up to amplification and dissociation curves and statistical analysis was done based on the optimized conditions.Results Through 25 tests the best real-time quantitative PCR conditions for ICAM-1 are anneal temperature at 61 ℃,0.4?g of cDNA and concentration of 150?mol/L each primer in the final reaction.There are no interaction among above factors.Conclusion Orthogonal design is a feasible,quick and economical method to optimize real-time quantitative PCR conditions.
