Am planning to detect lncRNA in serum of colon cancer patients. The plan is to test the samples in predispensed primer pcr array wells (Qiagen). Then do validation studies of individual qpcr assays.
This is the part i dont understand. Why do we have to do individual qpcr assays?
Am a doctor and just entering into the research field. All this is very new to me. Any advice would be appreciated
Dear Nab
I imagine that you will be using an assay to investigate the presence of total long coding RNA from your target samples. Once you've done this, you'll be comparing to a control of sorts to identify those which have differential expression at significant levels. Once you've identified these you will need to do quantitative qPCR to validate the differential expression for each as with long coding RNA candidates prioritised as there are a number of factors that may influence the yield obtained independent of the hypothesis you're testing. That said, regulation may be occurring at genomic, transcriptomic or proteomic levels and therefore you need to confirm whether the long coding RNA differences you observe correlate to the actual gene expression levels they are supposed to be responsible for.
I hope this helps.
Regards,
Nathaniel
Mcgregor explain the validation. If you have already small sample, you adjust my theory "the k-fold cross validation for small sample method" from downloading my papers. I validate all result by this method except for feature selection of microarray datasets.
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