n Methods Useful in Genetics and Oncology
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Fig. 3A-D. Quantitative peR of IL-6. A Monitoring of entire SYBR Green I peR reaction. Fluorescence vs. cycle number plot of IL-1 ~ cDNA amplification of 10-10 5 copies of standard and
two unknown samples (black arrows). B Identification of log-linear cycles for quantification;
green line denotes crossing line. Log fluorescence vs. cycle number plot of 1L-6 cDNA amplification. No-template negative control (blue line) and 10-105 copies of IL-6 cDNA standard (1
copy=green; 10 copies=red; 10 2 copies=black; 10 3 copies=pink; 104copies=dark green;
10 5 copies=dark blue) and two unknown myocardial biopsy samples (x and y). C Standard curve
of known copy number standards for IL-6 (slope: -3.1; intercept: 33.94; error: 0.233; r: -0.99). D
Melting curve analysis of peR products from all reactions
Table 2. Quantification of cytokine mRNAs in myocardial biopsy samples
Sample X
Sample Y
Copy number
Absolute Relative
Absolute
Relative
/1()6 ~-actin mRNA
I J()6 p-actin rnRNA
TNF-a
81
24
29
5
lL- l~
267
60
238
42
lL-6
688
200
297
52