miRNA Detection
BioLiqX HS miRNA qRT-PCR Kits for High Sensitivity miRNA Detection
• Pre-amplification step and negation of dilution effect ensure superior sensitivity
• Perfect for low copy number circulating miRNAs in biological fluids, such as plasma
• High specificity for any miRNA(s) of your choice; premade kit for organ and mestasis specific miRNAs
• Ideal for validating Small RNA-Seq results quantitatively
• Based on CATL technology: BioLiqX Small RNA-Seq results can be validated directly on the same libraries using BioLiqX HS miRNA qRT-PCR Assays
Workflow of the BioLiqX HS miRNA qRT-PCR Assays
The kits utilize a single-tube protocol for the conversion of all short RNA molecules into a preamplified cDNA library by adding certain reagents and enzymes to the RNA sample in a sequential manner. Subsequently, the preamplified cDNA serves as a template for ultra-sensitive and low-background real-time PCR reactions with Green DNA Dye (a SYBR® Green analog) and miRNA-specific primers. High specificity and sensitivity of BioLiqX HS miRNA Assays are achieved by the optimal concentration of nucleotides, mineral salts, Green DNA Dye and Hot Start Taq DNA Polymerase in the final qPCR reaction. All reactions can be set up at room temperature without the risk of non-specific amplification. The BioLiqX HS miRNA Assays are ideal for detecting low copy number circulating miRNAs in biological fluids but can be also applied to any other sample. The whole procedure can be completed within approximately 8 hours and requires typically a hands-on time between 30-60 minutes depending on the number of samples.
The BioLiqX HS miRNA Assays allow detection of very short cDNA fragments including truncated microRNAs and isomiRs. In addition, pre-amplified libraries generated by the protocol can be used for the simultaneous qPCR analysis of various other cell-free RNA fragments using custom primer pairs.
Superior Assay Performance of BioLiqX HS miRNA qRT-PCR Assays:
BioLiqX HS miRNA Assays have superior sensitivity compared to preamplification-free protocols mainly due to negating RNA and cDNA dilution effect during RT-qPCR and due to the implemented amplification step. The figure below shows the superior sensitivity of the BioLiqX HS miRNA hsa-miR-16, hsa-miR-21 and cel-miR-39 qRT-PCR assays compared to the corresponding preamplification-free protocols from two competing research kits:
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