Inter-laboratory collaborative trial of real-time PCR method for the relative quantitation of horse DNA and pork DNA in raw and processed beef DNA: phase 1
Inter-laboratory collaborative trial of real-time PCR method phase 1: Appendices
Appendices including tables and images for the report.
Appendix 1 Table A1
Spectrophotometer (NanoDropTM 1000) data for beef, horse and pork in raw and processed formats across the various % w/w combinations. The horse and pork % w/w/ combinations were in a matrix of beef. The data relate to individual extracts prior to pooling, absorbance at 260 and 280 nm.
| Sample matrix | A260 | A280 | A260/A280 | A260/A230 | ng/µL |
|---|---|---|---|---|---|
| 100% raw Beef extract 1 | 3.23 | 1.60 | 2.02 | 1.98 | 161.33 |
| 100% raw Beef extract 2 | 3.15 | 1.54 | 2.04 | 2.06 | 157.32 |
| 100% raw Beef extract 3 | 2.87 | 1.42 | 2.03 | 1.97 | 143.70 |
| 100% raw Horse extract 1 | 2.21 | 1.12 | 1.98 | 1.99 | 110.69 |
| 100% raw Horse extract 2 | 2.75 | 1.39 | 1.97 | 2.06 | 137.32 |
| 100% raw Horse extract 3 | 2.24 | 1.13 | 1.98 | 1.77 | 112.02 |
| 100% raw Pork extract 1 | 4.8 | 2.39 | 2.01 | 2.03 | 240.08 |
| 100% raw Pork extract 2 | 4.6 | 2.28 | 2.02 | 2.13 | 230.25 |
| 100% raw Pork extract 3 | 4.31 | 2.16 | 2.00 | 2.00 | 215.73 |
| 10% raw Pork extract 1 | 3.68 | 1.88 | 1.96 | 2.24 | 183.89 |
| 10% raw Pork extract 2 | 3.44 | 1.74 | 1.98 | 2.24 | 172.04 |
| 10% raw Pork extract 3 | 3.12 | 1.58 | 1.98 | 2.22 | 156.01 |
| 3% raw Pork extract 1 | 3.41 | 1.74 | 1.96 | 2.17 | 170.25 |
| 3% raw Pork extract 2 | 4.35 | 2.21 | 1.97 | 2.19 | 217.42 |
| 3% raw Pork extract 3 | 3.37 | 1.71 | 1.97 | 2.19 | 168.72 |
| 1% raw Pork extract 1 | 2.51 | 1.27 | 1.97 | 2.13 | 125.53 |
| 1% raw Pork extract 2 | 1.93 | 0.97 | 2 | 2.03 | 96.68 |
| 1% raw Pork extract 3 | 2.04 | 1.02 | 2.01 | 2.18 | 102.10 |
| 0.5% raw Pork extract 1 | 4.59 | 2.37 | 1.93 | 1.87 | 229.28 |
| 0.5% raw Pork extract 2 | 4.4 | 2.24 | 1.97 | 2.19 | 220.11 |
| 0.5% raw Pork extract 3 | 3.29 | 1.68 | 1.95 | 2.12 | 164.40 |
| 0.1% raw Pork extract 1 | 3.15 | 1.63 | 1.93 | 1.83 | 157.40 |
| 0.1% raw Pork extract 2 | 2.82 | 1.43 | 1.97 | 2.13 | 140.83 |
| 0.1% raw Pork extract 3 | 2.63 | 1.34 | 1.96 | 2.05 | 131.30 |
| 10% processed Pork extract 1 | 1.25 | 0.71 | 1.76 | 0.96 | 62.40 |
| 10% processed Pork extract 2 | 0.81 | 0.39 | 2.05 | 1.96 | 40.45 |
| 10% processed Pork extract 3 | 0.76 | 0.38 | 2.01 | 1.9 | 38.15 |
| 3% processed Pork extract 1 | 0.87 | 0.46 | 1.91 | 1.48 | 43.67 |
| 3% processed Pork extract 2 | 0.91 | 0.47 | 1.92 | 1.65 | 45.37 |
| 3% processed Pork extract 3 | 0.86 | 0.44 | 1.97 | 1.88 | 43.19 |
| 1% processed Pork extract 1 | 0.93 | 0.48 | 1.94 | 2.36 | 46.32 |
| 1% processed Pork extract 2 | 0.9 | 0.47 | 1.91 | 2.09 | 45.24 |
| 1% processed Pork extract 3 | 0.84 | 0.44 | 1.9 | 2.32 | 42.21 |
| 0.5% processed Pork extract 1 | 1.29 | 0.68 | 1.91 | 2.22 | 64.73 |
| 0.5% processed Pork extract 2 | 1.23 | 0.65 | 1.91 | 2.04 | 61.73 |
| 0.5% processed Pork extract 3 | 0.94 | 0.49 | 1.92 | 1.95 | 47.14 |
| 0.1% processed Pork extract 1 | 1.33 | 0.72 | 1.83 | 1.51 | 66.25 |
| 0.1% processed Pork extract 2 | 1.1 | 0.59 | 1.86 | 1.71 | 54.93 |
| 0.1% processed Pork extract 3 | 1.32 | 0.7 | 1.9 | 2.1 | 66.23 |
| 10% raw Horse extract 1 | 4 | 2.07 | 1.93 | 2.14 | 199.83 |
| 10% raw Horse extract 2 | 3.7 | 1.91 | 1.93 | 2.15 | 185.10 |
| 10% raw Horse extract 3 | 3.97 | 2.04 | 1.95 | 2.16 | 198.31 |
| 3% raw Horse extract 1 | 3.92 | 2.03 | 1.93 | 2.16 | 195.80 |
| 3% raw Horse extract 2 | 2.95 | 1.52 | 1.93 | 2.1 | 147.43 |
| 3% raw Horse extract 3 | 4.16 | 2.14 | 1.94 | 2.18 | 208.20 |
| 1% raw Horse extract 1 | 3.12 | 1.61 | 1.94 | 1.93 | 156.18 |
| 1% raw Horse extract 2 | 3.56 | 1.81 | 1.97 | 2.2 | 178.06 |
| 1% raw Horse extract 3 | 3.16 | 1.64 | 1.93 | 1.81 | 158.20 |
| 0.5% raw Horse extract 1 | 3.97 | 2.04 | 1.95 | 2.06 | 198.32 |
| 0.5% raw Horse extract 2 | 3.34 | 1.71 | 1.95 | 2.11 | 166.86 |
| 0.5% raw Horse extract 3 | 3.36 | 1.72 | 1.96 | 2.23 | 167.92 |
| 0.1% raw Horse extract 1 | 4.56 | 2.34 | 1.95 | 2.19 | 227.99 |
| 0.1% raw Horse extract 2 | 3.3 | 1.7 | 1.95 | 2.21 | 165.07 |
| 0.1% raw Horse extract 3 | 4.49 | 2.32 | 1.94 | 2 | 224.74 |
| 10% processed horse extract 1 | 1.19 | 0.62 | 1.93 | 1.81 | 59.40 |
| 10% processed horse extract 2 | 1.27 | 0.66 | 1.93 | 1.88 | 63.37 |
| 10% processed horse extract 3 | 1.35 | 0.71 | 1.91 | 1.73 | 67.49 |
| 3% processed horse extract 1 | 0.94 | 0.49 | 1.91 | 1.73 | 47.22 |
| 3% processed horse extract 2 | 1.05 | 0.53 | 1.98 | 1.93 | 52.35 |
| 3% processed horse extract 3 | 1.23 | 0.62 | 1.98 | 1.93 | 61.67 |
| 1% processed horse extract 1 | 1.29 | 0.76 | 1.7 | 0.79 | 64.41 |
| 1% processed horse extract 2 | 1.58 | 0.93 | 1.69 | 0.81 | 79.08 |
| 1% processed horse extract 3 | 1.45 | 0.86 | 1.68 | 0.77 | 72.51 |
| 0.5% processed horse extract 1 | 1.53 | 0.84 | 1.81 | 1.78 | 76.58 |
| 0.5% processed horse extract 2 | 1.54 | 0.84 | 1.84 | 1.91 | 77.05 |
| 0.5% processed horse extract 3 | 0.9 | 0.5 | 1.81 | 1.78 | 45.24 |
| 0.1% processed horse extract 1 | 1.43 | 0.77 | 1.85 | 2.21 | 71.45 |
| 0.1% processed horse extract 2 | 1.53 | 0.83 | 1.85 | 2.24 | 76.48 |
| 0.1% processed horse extract 3 | 1.53 | 0.83 | 1.84 | 2.22 | 76.73 |
Appendix II: Authentication of meat species by Sanger sequencing, alignment figures
Figure A1: Alignment figure for pork forward and reverse cytochrome B sequences against the pork reference sequence (NC_000845).
Figure A2: Alignment figure for horse forward and reverse cytochrome B sequences against the horse reference sequence (D82932).
Figure A3: Alignment figure for beef forward and reverse cytochrome B sequences against the beef reference sequence (D34635).
