Low volume amplification in single cell separation and inspection.
- Author:
Jun-Ping HAN
1
;
Cai-Xia LI
;
Hong YAN
;
Dian ZHU
;
Gen-Ping LI
;
Lan HU
Author Information
1. Chinese People's Public Security University, Beijing 100038, China.
- Publication Type:Journal Article
- MeSH:
Alleles;
Cell Separation/methods*;
DNA/genetics*;
DNA Fingerprinting/methods*;
Endopeptidase K/administration & dosage*;
Enzymes/administration & dosage*;
Epithelial Cells/cytology*;
Feasibility Studies;
Forensic Genetics;
Genotype;
Humans;
Mouth Mucosa/cytology*;
Nucleic Acid Amplification Techniques;
Polymerase Chain Reaction/methods*;
Sensitivity and Specificity
- From:
Journal of Forensic Medicine
2012;28(2):123-125
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish optimal amplification conditions with the application of 0.2 mL test tube in single cell separation and inspection.
METHODS:Oral epithelium cell suspension was prepared. Five or ten cells were collected with 0.2 mL test tube. Then DNA were amplified with Identifiler Plus kit in three different conditions in which the proteinase K addition, gold enzyme concentration in PCR reaction, and PCR reaction cycles were adjusted separately. Finally the detection rate, allelic dropout rate and artificial alleles were compared among the groups.
RESULTS:In these 3 different conditions: addition of proteinase K, addition of 0.4 microL gold enzyme in PCR reaction, and use of 32 cycles, the detection rate was higher and allelic dropout rate was lower than the other conditions.
CONCLUSION:In single cell separation and inspection, the usage of 0.2 mL test tube could be an effective supplement to chip-low volume amplification.
