Extraction of DNA Using Monoclonal Anti-DNA and Colloidal-gold Beads
- VernacularTitle:免疫胶体金法提取环境标本中细菌DNA技术
- Author:
Jie-Ping PEI
;
Jun HE
;
Hua TAN
;
Hong ZHU
;
Qing DUAN
;
- Publication Type:Journal Article
- Keywords:
Monoclonal anti-DNA,DNA extracting,PCR
- From:
Microbiology
1992;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
To developing a simple, rapid and sensitive immunoaffinity method for extracting DNA before amplification by the polymerase chain reaction(PCR). Using monoclonal anti-DNA bound to colloidal-gold beads(which called immunocolloidal-gold ) and extracting DNA from the specimen before amplification by PCR. The immunoaffinity method we use is extremely efficient in extracting DNA at low concentration, also, the beads carrying the DNA/anti-DNA complexes can be added directly to a PCR mixture without elution, and the presence of PCR inhibitors in specimens has no effect on amplification. When applied to serial dilution bacteria suspension, the detection of limit can be 100 CFU/mL, and the sensitivity of detecting artificial soil sample and milk sample were 101 and 100 CFU/mL. The immunoaffinity method described here can sever as a sensitive and practicable method for extracting DNA , particularly where samples have low concentrations of DNA or where the material is in poor condition.