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Blood, Vol. 113, Issue 20, 4914-4917, May 14, 2009
Deregulation of microRNA involved in hematopoiesis and the immune response in HTLV-I adult T-cell leukemia
Blood Bellon et al.
113: 4914
Supplemental materials for: Bellon et al
MicroRNA bioarrays: The mirVana miRNA Bioarrays platform was used (Ambion, Inc.) with a Molecular devices 4200AL scanner performed by Asuragen Inc (Austin, TX). The fluorescent signal (probes and local background) was extracted using GenePix Pro followed by one-way ANOVA (Analysis Of Variance) statistical testing. Stem loop quantitative real time RT-PCR: Reverse transcription, including no-template controls and RT minus controls, were run in duplicate. TaqMan Real-time PCR was performed on a 7900HT System (Applied Biosystems). All reactions were run in triplicate. Statistical analysis is provided as Supplemental 2. miR-24 expression was used as an internal control. For microRNAs, miR-125a, -132, -150, -155, -181a, and -223, values are normalized to T-cell control. For miRNA, miR-142-3p, values are normalized to PBMC control. Statistical analyses was performed as follows: Undetermined raw Ct values were set to 40. Median Ct replicate values were calculated, and each plate scaled to the negative plate median Ct value. ΔCt and ΔΔCt computation: The ΔCt values are computed independently for the “unknown sample” and the “calibrator” assay (mir-24) as a difference in the Average Ct values. The ΔΔCt values indicate the gene expression as a difference between the ΔCt for the tested “unknown sample” and the control sample miR-24. The fold changes between genes are expressed as 2-(ΔΔCt) values or the negative inverse of that equation, if that value is less than 1. All amplifications are performed on an ABI 7900HT real-time thermocycler and data analyzed with the provided software. ND = not detected (not expressed). St. Dev = standard deviation.
Files in this Data Supplement:
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