• ID3EAL Spike-in Solution

    Variability in sample processing steps (such as RNA isolation) could lead to differences in the quality and quantity of the RNA extracted resulting in inaccurate data interpretation downstream. The best way to monitor and normalize for these batch-to-batch differences is to incorporate standard spike-in controls into the workflow.

    MiRXES ID3EAL Spike-in kit consists of uniquely designed synthetic small RNAs (~22 nt) with sequences distinct from endogenous miRNAs, their corresponding RT-qPCR primers, and an optimized master mix. The kit had been extensively tested and shown to be compatible with various isolation methods, including phenol/chloroform-based, phenol-free, membrane, bead and precipitation methods.
    Spike-in controls may be incorporated during the RNA isolation and/or Reverse Transcription steps of the RNA-to-Ct workflow; Spike-ins added in the isolation process would capture workflow information from isolation until qPCR while Spike-ins added during cDNA synthesis would capture workflow information from the cDNA synthesis step onwards.
  • Verification of Workflow

    Table illustrating possible scenarios leading to Ct value greater than 30 for Spike-in controls.

    The efficiency of the various steps could be determined easily by the consistency of the Ct values obtained; Ct value higher than 30 indicates poor isolation and/or cDNA synthesis efficiency.