miRNA Profiling: Save Time and Sample Using Real-Time PCR
|Streamlined workflow enables miRNA profiling using real-time RT-PCR as an alternative to microarrays: |
Using Real-time-PCR for miRNA Profiling
Figure 1. Save 45 Steps Using Multiplex RT Pools Workflow. Multiplex RT pools can be used in place of individual MicroRNA RT primers (singleplex RT) to increase throughput for miRNA analysis, decreasing the workflow by 45 steps.This makes miRNA expression profiling possible using TaqMan® chemistry, while enabling increased speed and additional sensitivity over array expression profiling methods. Note that Multiplex RT pools can be used with TaqMan Low Density Arrays for a further enhancement to the workflow.
Only 8 RT reactions are required to obtain coverage of 365 human miRNAs and two endogenous control genes using the complete set of Multiplex RT pools. This is in contrast to the 300–400 individual miRNA RT reactions that would otherwise be required, and results in tremendous time and cost savings.
Faster Profiling with Less Sample: Just 3 hr and 200 ng for a Human miRNA Profile
|Product Configuration||Recommended Range (per Rxn)||No. RT Rxns||Total RNA Input|
|TaqMan||1-10 ng/Rxn||325||325 ng - 3.25 µg|
|Multiplext RT||20-400 ng/Rxn||8||160 ng - 3.2 µg|
Figure 2. Sample Input Requirements for miRNA Screening with TaqMan® MicroRNA Assays (365 miRNA targets + 2 endogenous control genes).