miScript PCR Starter Kit
For setup and optimization of miRNA quantification experiments
For researchers starting out in the field of miRNA research, the miScript PCR Starter Kit provides all the necessary components for set-up of miRNA quantification experiments. The kit includes reagents for reverse transcription and real-time PCR, a miScript PCR Control for RNU6-2 for normalization, and a miScript Primer Assay for detection of human miR-15a.
Performance
miR-15a is highly expressed in human tissues and cells, making it ideal for use as a positive control. The miScript PCR Starter Kit enables accurate, linear detection of miR-15a in positive control experiments for setup and optimization of miRNA quantification (see figure Accurate, linear detection of miR-15a).
Principle
The miScript PCR System enables easy miRNA quantification and profiling using SYBR® Green-based real-time PCR. The miScript PCR System covers all the steps of conversion of RNA into cDNA to detection of miRNAs in SYBR® Green-based real-time PCR. The miScript PCR Starter Kit combines the key components of the miScript PCR System in a single kit, allowing the user to establish miR-15a quantification and normalization of results, before proceeding to miRNA quantification in their field of interest.
Procedure
cDNA prepared in a reverse transcription reaction using miScript HiSpec Buffer or miScript HiFlex Buffer serves as the template for real-time PCR. Real-time PCR is performed using a human miR-15a-specific miScript Primer Assay (forward primer) and the miScript Universal Primer (reverse primer) with QuantiTect SYBR Green PCR Master Mix. For normalization, a parallel reaction is performed using the human RNU6-2 (RNU6B) miScript Primer Assay and the miScript Universal Primer with QuantiTect SYBR Green PCR Master Mix.
Applications
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