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miScript PCR Starter Kit

For setup and optimization of miRNA quantification experiments

  • Straightforward setup of miRNA quantification experiments
  • Easy monitoring and optimization of miRNA quantification
  • Reliable positive and normalization controls

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.

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Cat No./ID: 218193
miScript PCR Starter Kit (80)
For 10 x 20 μl RT reactions and 80 x 25 μl PCRs; miScript Reverse Transcriptase Mix, 10x miScript Nucleics Mix, 5x miScript HiSpec Buffer, 5x miScript HiFlex Buffer, 2x QuantiTect SYBR Green PCR Master Mix, 10x miScript Universal Primer, Human RNU6B (RNU6-2) miScript Primer Assay, Human miR-15a miScript Primer Assay, RNase-Free Water
The miScript PCR Starter Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

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Accurate, linear detection of miR-15a.

RNA was purified from HeLa S3 cells. Using the miScript PCR Starter Kit, cDNA was generated and a range of cDNA amounts from 10 pg to 10 ng was used in real-time PCR to detect miR-15a. Real-time PCR was performed in the Rotor-Gene Q cycler. The resultant CT values decreased linearly with increasing cDNA amount, indicating quantitative detection from a wide range of template amounts.
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
  • Set up of miRNA quantification experiments
  • miRNA control experiments

Product Resources

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Kit Handbooks (1)
For real-time PCR analysis of microRNA using SYBR Green detection
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