Blocking oligo--a novel approach for improving chip-based DNA hybridization efficiency.
For most of the commonly used DNA chips, the probes are usually single-stranded oligonucleotides and the targets are double-stranded DNAs (dsDNAs). Only one strand of the DNA serves as the target while the other competes with the probes immobilized on the chip for the target and therefore is regarded as the interfering strand. In this report, a novel technique was developed for improving the hybridization efficiency on DNA chips by using blocking oligos, which is complimentary to the target interfering strand to reduce the influence of the interfering strand. The hybridization efficiency of dsDNA was much lower than that of single-stranded DNA (ssDNA) when synthesized DNA targets were tested on the DNA chip. Blocking oligos can improve the hybridization efficiency of dsDNA to about 2/3 that of ssDNA. Blocking oligos have also been applied to PCR products of different lengths for hybridization. The hybridization efficiency with blocking oligos is about three times higher than that without blocking oligos. We have tested PCR products of 1054 and 435 bp using our blocking procedure, and the results are consistent.
Tao
Sheng-ce
SC
Department of Biological Sciences and Biotechnology, Tsinghua University, Haidian District, 100084, Beijing, People's Republic of China.
Gao
Hua-fang
HF
Cao
Fei
F
Ma
Xue-mei
XM
Cheng
Jing
J
D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004277:DNA, Single-Stranded / Q000032:analysis; D005456:Fluorescent Dyes; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes
D015342:DNA Probes; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D004247:DNA