1. Which is the best kit for the isolation of microRNAs from serum samples?
2. Which microarray platform gives the best results for high-throughput miRNA expression analysis?
3. How much starting material (serum) in terms of quantity (amount) will be enough to give miRNA for microarray experiments.
Hi. With regards to your first question, I found this paper quite useful: http://www.ncbi.nlm.nih.gov/pubmed/22982505
I compares three commercially available kits for miRNA isolation from clinical serum samples using 200 microlitres of starting material. I hope it helps.
Hi
I agree with José about the first part of the question. The Exiqon kit for biofluids is nice, reliable and relatively cost-effective.
About the second question, I will not recommend you to use microarrays for miRNA profiling. It would be better to use pPCR panels for screening. The reason is related with the size of miRNAs, which will bias your results towards the most abundant miRNAs. Microarrays are great for detecting things with relatively high expression levels, but because of the size of miRNAs, you will lose a lot of information about the less expressed ones. Moreover, in many conditions the less expressed miRNAs are the most relevant since they can be subjected to more dramatic changes.
Hope it helps. Best
Paco
Thanks Jose.....The paper puts up nicely the comparative assessment of three kits and QIAGEN kit comes out to be the winner....I was also thinking about going with the same but this paper now made my task easier.
Thanks Francisco....Sensitivity is an issue with microarray. However, can we use qPCR panels to quantify the expression of all mature miRNAs? As far as I know, we can only profile a limited set of miRNAs with qPCR while with microarray, we can profile around 2000 mature miRNA sequences...Please convey ur opinion....
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