Influences of cycle index and volume on sensitivity of DNA detection.
- Author:
Lu ZHANG
1
;
Bao-Jie WANG
;
Mei DING
;
Zi-Qing LIN
;
Hao PANG
;
Jia-Xin XING
;
Jin-Feng XUAN
Author Information
1. School of Forensic Medicine, China Medical University, Shenyang 110001, China. zhangluzhaoyang@sohu.com
- Publication Type:Journal Article
- MeSH:
DNA/genetics*;
DNA Fingerprinting/methods*;
Feasibility Studies;
Forensic Genetics/methods*;
Humans;
Limit of Detection;
Male;
Polymerase Chain Reaction/methods*;
Semen/chemistry*;
Sensitivity and Specificity;
Tandem Repeat Sequences
- From:
Journal of Forensic Medicine
2013;29(2):125-126
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the feasibility of improving the sensitivity of DNA detection by increasing the PCR cycle index and decreasing the volume of amplifying system.
METHODS:The DNA of semen were collected from 10 healthy irrelevant volunteers, and were quantified to 50, 40, 30, 25, 20, 15, 10 pg/microL, separately. All samples were then amplified in 10, 5, 3 microL volume and at 28, 30, 32, 34, 36 cycles, respectively. 3130 genetic analyzer was used to detect 15 autosomal STR loci.
RESULTS:Under the situation of 28 cycles and 3 microL volume, samples which achieved > 40 pg/microL could be correctly typed. Under the situation of 10, 5, 3 microL volume, samples which achieved > 20 pg/microL could be correctly typed at 34 cycles. When increasing the index to 36 cycles, they could not be correctly typed because of the non-specific band.
CONCLUSION:DNA detecting sensitivity can be improved to a certain extent by increasing the cycle index and decreasing the volume of amplifying system.
