I'm currently working on the isolation of miRNA from human serum samples. Using the column based protocol, I have been able to isolate around 5ng/ul of miRNA, but the quality of RNA (260/280) using a NanoDrop, and the ratio was 1.3 and I understand this is may be insufficient to proceed to Array study to determine differentially expressed miRNA from clinical samples.
We isolated miRNA from human serum samples by using miRNeasy Mini Kit for tissue/cell (Qiagen,Cat. No: 217004) and we also obtained 1-5 ng/ul of miRNA and it was very low to progress with array. However, we converted miRNA into cDNA and obtained about 500 ng/ul. Then, we studied with real time for miR expression analysis, it worked very efficiently. From this point, even if we used tissue/cell kit instead of serum based kit and measurements were very low, it worked in Real Time. So your samples would work on array, I think.
Dear Roberto
Running microRNAs from serum or any body fluids is always challenging and in our opinion arrays aren't the best solution.
At NXT-Dx, we offer miRNA expression profiling using the latest in qPCR technology. We have compared qPCR,with arrays and sequencing techniques, especially in body fluids such as serum. The qPCR approach clearly comes forward as the optimal solution. On top of this we specialize in low-input approaches with this technique, so are very capable in delivering high quality data from serum samples. We can also take care of the RNA isolation from your serum samples before starting the
If you are still planning to run this experiment and would be interested in learning more about our approach, please send me an email on [email protected].
Maarten Braspenning
NXT-Dx
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