The miRNA array enables exploration of the regulatory mechanisms mediated by miRNAs and their importance in cancer and other diseases. The array is also suitable for formalin-fixed, paraffin-embedded (FFPE) tissue sample analysis. The analysis can be performed as single arrays or in 96 array plate format
|Research Objectives||Measurement of small non-coding RNA transcripts involved in gene regulation|
Low molecular weight molecules have been shown to be involved in a number of biological events including mRNA degradation, transcriptional gene silencing and translational repression. A specific assay is used that enables the labelling of total RNA and low molecular weight RNA. An ELOSA (Enzyme Linked Oligosorbent Assay) QC step is carried out to confirm the success of the biotin labelling process. Positive and negative controls are included to monitor assay performance. These arrays also include quality control probes to assess successful hybridisation to the arrays.
The number of samples to be analysed is dependent on the nature of the experimental design. It is advisable to have at least 3 replicates however the actual optimal number depends on the samples to be investigated and variance inherent in the experimental system. Following QC steps, the samples undergo Poly (A) tailing followed by a ligation step to generate biotin-labelled RNA. An ELOSA QC step is performed prior to sample hybridisation to determine successful labelling. The prepared targets are hybridized to the arrays for 16-18 hours, washed and stained using the GeneChip Fluidics Station 450 and scanned using the GeneChip® Scanner 3000 7G. The initial data QC analysis is carried out to determine the quality of the data using Affymetrix Expression Console.
An analytical report is provided which includes the QC results for the ELOSA performed prior to hybridisation. This is accompanied by a separate data QC report and the raw data files.
- RNA Quantity: 10µg
- RNA Concentration: 100 ng/µl