[Preliminary exploration of the influence factors of degenerate oligonucleotide primered PCR of genome DNA].
OBJECTIVE
To explore the influence factors of the degenerate oligonucleotide primered PCR(DOP-PCR).


METHODS
Genome DNA template from the mouse single oocyte or liver tissue were used to perform DOP-PCR. DOP-PCR was carried out with templates of different origin, different gradient dilution, with or without low melting point gel purified to wipe off the small fragment that might interfere with the following analysis, and then PCR of gene FTCD and CBS were carried out to evaluate the influence of these factors on the amplification efficiency and specificity.


RESULTS
Compared with genome DNA template from mouse liver, the template from single oocyte had the same efficiency and specificity but a minor yield and different gradient dilution of DNA template had no effect on the efficiency and specificity. Furthermore, there was a higher specificity in the low melting point gel-purified DOP-PCR product than in untreated ones.


CONCLUSIONS
We have got a satisfactory result and increased specificity from DOP-PCR product purified with the low melting point gel. Single oocyte of mice could be used for further investigation of special genes detection by DOP-PCR and of an optimization in the yield of the products.
Yang
Li-jun
LJ
Department of Paediatric, Peking University People's Hospital, Beijing 100044, China.
Yu
Wei-dong
WD
Liang
Rong
R
Shang
Mei
M
Guo
Jing-zhu
JZ
D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D016678:Genome; D005838:Genotype; D051379:Mice; D008807:Mice, Inbred BALB C; D009841:Oligonucleotides / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015203:Reproducibility of Results; D018401:Sample Size; D017422:Sequence Analysis, DNA; D013698:Templates, Genetic
D017931:DNA Primers; D009841:Oligonucleotides