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Chemical Metrology and Biometry Department National Institute of Metrology (Thailand)

CCQM P103.1 “ Measurement of multiple RNA transcripts”. Duangkamol Viroonudomphol Chaiwat Prawettongsopon. Chemical Metrology and Biometry Department National Institute of Metrology (Thailand). Real-time PCR . Instrument: ABI 7500 Real Time PCR System

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Chemical Metrology and Biometry Department National Institute of Metrology (Thailand)

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  1. CCQM P103.1 “Measurement of multiple RNA transcripts” DuangkamolViroonudomphol ChaiwatPrawettongsopon Chemical Metrology and Biometry Department National Institute of Metrology (Thailand)

  2. Real-time PCR Instrument: ABI 7500 Real Time PCR System RT-qPCR kit: AgPath-ID RT-PCR Reagents kit (ABI) Reaction: 10 µl, RNA sample = 1 µl/reaction Analysis software: 7500 Software V 2.0.6

  3. ERCC measurement: Absolute quantification

  4. Optional GOIs: Relative quantification • Based on a doubling of template with each PCR cycle 2 -∆Cq(U1-U2) • Normalised with CASC3 GOIs/CASC3

  5. Source of uncertainty budgets ERCCs: Regression from the calibrantion, Precision, Pipetting bias GOIs: Precision, Pipetting bias Relative expression ratio (a) = U1/U2 = b/c ua = a √ (ub/b)2 + (uc/c)2

  6. THANK YOU

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