I have been wondering what minimal amount of RNA may be successfully used for TaqMan Array MicroRNA cards as the manufacturer recommends 350 – 1000 ng to be used without pre-amplification, however, in low-concentration samples this is not often available. Thank you for your suggestions.
Hi Luděk,
I've worked with 2ng/ul of RNA extraction (which contains the miRNAs), with TaqMan microRNA Assays (Applied biosystems).
I have used 1 ng total RNA into the LifeTechnologies/Thermo MegaPlex miRNA system with no problems.
Hi Ludek,
Alternatively you could make a preamplification step. Life tech. sells a preamplification primer mix for human or mouse. If you use it do not forget to premp. all your samples to correct for posible biases.
Hope this helps
Estanis
HI!!
I recommend you to use pre-amplification step if you have 1-2 ng.
search for applied biosystem part number 4384556 for a quick reference of how to make your own preamplification pool for the miRNAs of interest.
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