[PCR in combination with silver as a method for the determination of apolipoprotein E alleles].
OBJECTIVE
To present a rapid, sensitive and safe procedure for the determination of human apolipoprotein E alleles.


METHODS
Genomic DNA was extracted from 0.5ml whole blood by a non-phenol protocol. After PCR and restriction enzyme digestion,short DNA fragments were detected by a simplified silver staining method. This technique was used to assay apolipoprotein E alleles in 19 patients with sporadic Alzheimer's disease and 41 normal aged persons.


RESULTS
The short DNA fragments were visualized clearly on polyacrylamide gel processed by silver staining. It took 24 hours from DNA extraction to the final result. The A4 allele frequency in patients with sporadic Alzheimer's disease was 0.29, much higher than that in control group (0.02, P<0.001).


CONCLUSIONS
PCR in combination with silver staining can satisfy the need for high-resolution and high-efficiency in the determination of apolipoprotein E alleles and can be used as a routine procedure in clinical and epidemiological investigations.
Institute of Geriatrics, PLA General Hospital, Beijing 100853 P. R. China. zaqqhw@ihw.com.cn
A
A
Zhang
Z
Z
Han
L
L
Wang
G
G
Tao
D000483:Alleles; D000544:Alzheimer Disease / Q000235:genetics; D001057:Apolipoproteins E / Q000235:genetics*; D004396:Coloring Agents; D004247:DNA / Q000032:analysis; D006801:Humans; D016133:Polymerase Chain Reaction; D012834:Silver
D001057:Apolipoproteins E; D004396:Coloring Agents; D012834:Silver; D004247:DNA