A modified hybridization protocol for low-density diagnostic DNA microarrays.
To promote the application of DNA microarrays for clinical diagnosis, the problems of cross-hybridization and low signal intensity in the hybridization processes has been addressed. We tested a new hybridization protocol for low-density diagnostic DNA microarrays, by skipping the purification step during sample labeling, while elevating the hybridization temperature from 42 degrees C to 52 degrees C, adding a step of distilled water rinsing immediately after hybridization and before the low stringency washing steps. It was found that the modified hybridization protocol works well in our study, which increased detection sensitivity and eliminated nonspecific signals.
Huang
Hai
H
Institute of Molecular Biology, First Military Medical University, Guangzhou, 510515, China.
Ma
Wen Li
WL
Wang
Hong Min
HM
Zhang
Bao
B
Wu
Qing Hua
QH
Zheng
Wen Ling
WL
D004269:DNA, Bacterial / Q000032:analysis*; D006358:Hot Temperature; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D015010:Yersinia pestis / Q000235:genetics / Q000302:isolation & purification
D004269:DNA, Bacterial