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We used a splicing microarray analysis to obtain alternative splicing events (ASEs) showing a significant change between MCF7 and MCF10.
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In this work, we used a splicing-sensitive microarray technology to investigate changes in pre-mRNA splicing that are associated to the formation of macroscopic metastases.
Using an alternative splicing microarray platform Pan and co-workers found that most of the PTC-containing transcripts were low in abundance across examined tissues, and this low abundance was independent of NMD function [ 41], arguing against the regulation function of NMD.
Candidate exons for alternative splicing were detected using a Splicing Index and the MIDAS algorithm [ 11].
Splice sites were predicted using a splice site finder program [ 34].
Any mutations that were believed to have an affect on splicing were analysed using a splice site prediction programme (http://fruitfly.org/seq_tools/splice.html).html
To experimentally test for alternative splicing caused by novel SNPs we used a minigene splicing assay.
This type of approach has already been shown to distinguish different alternative splice forms when used to design splicing microarrays (Johnson et al., 2003).
The trans-splicing technology uses a trans-splicing molecule that "tricks" the spliceosome into using it as a substrate for splicing.
Using a technique called microarray — which involves a "gene chip," the size of a saltine, that has embedded on it parts of the DNA sequences for thousands of genes — Karumanchi began to evaluate genes that are active in the placenta.
Finally, fluorescence images were captured using a microarray scanner (Axon).
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