[Investigation of genotyping HLA DRB1 gene using oligoneucleotide arrays].
We have developed a method performed on an oligoneucleotide array for genotyping HLA DRB1. The routine method Phenol-chloroform was used to extract genome DNA of standard samples. A pair of sense and antisense primers were designed according to the sequence of DRB1 exon2, then the primers and the Cy5-dCTP were used in the following PCR, thus the PCR products were labelled with Cy5. Many genotyping probes which were immobilized on the DNA Microarray made by APS-PDC method were designed. The labelled PCR products were hybridized with them, the signals were sanned by sanner and analyzed by Imagene software. We have genotyped 33 standard samples which have 12 DRB1 subtypes. The experimental results showed that the arrays we made and the method we used are accurate and sensitive. This proved that the DNA Microarray technique is good for DRB1 genotyping. Compared with PCR-SSP and PCR-SSO methods, the genotyping chip method is more intuitionistic and has the advantage of integration.
Chen
J N
JN
National Key Laboratory of Genetic Engineering, Institute of Genetics of Fudan University, Shanghai 200433, China.
Li
Y
Y
Li
Y L
YL
Qin
H Y
HY
Li
R Y
RY
Cao
H M
HM
Xie
Y
Y
Mao
Y M
YM
D005838:Genotype; D006684:HLA-DR Antigens / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction
D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains