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Sixth, our outcome will be measured by self-report, without biological verification.
For initial biological verification we focused on genes with established prominence-specific expression patterns such as Hand2 (GeneID: 15111), Lhx8 (GeneID: 16875), Elavl4 (GeneID: 15572) and members of the Dlx family [24], [25], [26], [27].
First, we had only self-reported data on substance use with no biological verification available.
Quantitative real time PCR was used for both technical (microarray) and biological verification.
Additional biological verification and characterization of these kinotypic differences will be the subject of a subsequent study.
Using new samples allows biological verification of the reproducibility of expression data (Additional file 6: Figure S1).
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Algorithms can never replace biological verifications, but should always be used to guide experimental design.
The proposed method can be used as a complementary tool besides baseline regression techniques, to provide a richer and a more promising list of candidate molecular predictors for further biological verifications.
Some of the AIMs identified in this study may be limited by the small to moderate number of samples in the HapMap II project; therefore, the generality of the identified AIMs should be further examined by using more independent samples and confirmed by biological verifications such as real-time reverse-transcription polymerase chain reaction before the AIMs applying to practical studies.
Third, the eQTL identified in this study might be limited by the small to moderate number of samples and ethnic populations available in the HapMapII project; therefore, the results should be further examined using additional samples from additional populations and require increased biological verifications.
Nevertheless, most of the countries that took part in the seven years of negotiations on biological weapons verification appeared resigned to the American withdrawal from the agreement.
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