RT-PCR report: experiment test 1

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RT-PCR report: experiment test 1 Sample description: Sample isolation date Lysate prep date cell type specimen age passage culture time O2 (%) equipment Adeno Flox operator # year month day 4 2014 1 19 29 chondrocyte 2 days 10 days 21 binder LM/CM 5 6 RNA quantification after DNase treatment: RNA integrity: ARN sample # [RNA] (ng/mL) Vol. eq. 500ng RNA (mL) Vol. eq. 1mg RNA (mL) Vol .eq. 2 mg RNA (mL) sample 4 584.2 0.9 1.7 3.4 sample 5 543.3 1.8 3.7 sample 6 541.1 RT-PCR summary: Summary of your RT-PCR experiment RNA input (mg) RT vol. (µl) Dilution RT Vol Dil RT (µl) ng/well 1 20 50

RT-PCR report: experiment test 1 Plate map:

RT-PCR report: experiment test 1 Validation of the results: ACAN -RT amplification curves: +RT-RNA amplification curves: No amplification: VALID No amplification: VALID +RT amplification curves: +RT melting curves: Amplification for all the samples: VALID Single peak and homogenous Tm: VALID

RT-PCR report: experiment test 1 Validation of the results: COL2A1 -RT amplification curves: +RT-RNA amplification curves: Late amplification >>>+RT: VALID No amplification: VALID +RT amplification curves: +RT melting curves: Amplification for all the samples: VALID Single peak and homogenous Tm: VALID

RT-PCR report: experiment test 1 Validation of the results: GAPDH -RT amplification curves: +RT-RNA amplification curves: No amplification: VALID No amplification: VALID +RT amplification curves: +RT melting curves: Amplification for all the samples: VALID Single peak and homogenous Tm: VALID

RT-PCR report: experiment test 1 Validation of the results: 18S -RT amplification curves: +RT-RNA amplification curves: Late amplification >>> +RT!: ~VALID Late amplification >>> +RT!: ~VALID +RT amplification curves: +RT melting curves: Amplification for all the samples: VALID Single peak and homogenous Tm: VALID

RT-PCR report: experiment test 1 Genes of reference distribution GAPDH 18S Ct Ct SEM = 0.1399 SEM = 0.1722 In these experimental conditions, GAPDH is more stable than 18S

RT-PCR report: experiment test 1 Analysis ACAN COL2A1 ACAN relative expression/GAPDH COL2A1 relative expression/GAPDH ACAN relative expression/18S COL2A1 relative expression/18S