New study just released compares three 60-mer oligonucleotide microarray hybridisation protocols for reproducibility, sensitivity, system noise, and accuracy
This study compares Agilent's standard procedure with two other hybridisation protocols, evaluating all three based on reproducibility, sensitivity, system noise, and accuracy.
The results indicate that while different hybridisation methods are available, use of Agilent's recommended procedures yields better microarray sensitivity and is likely to give more reproducible results with lower sample requirements Long oligonucleotide microarrays are becoming a widely used tool for the parallel expression analysis of thousands of genes.
To take full advantage of the benefits of long in situ oligonucleotide microarrays, the hybridisation conditions must be selected to yield the best possible results consistent with the goals of the experiment.
Frequently, research samples are limited and low abundance transcripts don't show significant signal above background.
The development of more sensitive techniques, which do not compromise the accuracy or precision of the data, enables larger amounts of significant data to be extracted from microarray experiments using 60-mer oligonucleotide microarrays. Prior to development of Agilent's in situ Hybridisation Reagent Kit with preformulated reagents, which improved ease-of-use and reproducibility, gene expression researchers had to use alternative hybridisation methods, many of which involved the use of toxic chemicals such as formaldehyde.
Although the data accuracy was comparable between the three different methods, the two alternative methods did not show the same level of sensitivity or reproducibility as Agilent's method.
For further information, request application note 'Comparison of hybridisation methods for Agilent's 60-mer in situ synthesised oligonucleotide microarrays', publication number 5988-5022EN.
This application note is available without charge from any Agilent sales office or its website (details above).
The in situ Hybridisation Reagent Kit, product number G2559A, is available by contacting your local Agilent Customer Call Centre or a local Agilent sales representative.
