Kit for detecting cow milk content in goat milk product and application of kit
A kit and goat milk technology, applied in the field of food safety, can solve the problems of low cell content and inability to judge negative results, etc., and achieve the effects of strong specificity, good reproducibility, and low detection limit
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Embodiment 1
[0047] Embodiment 1: experimental method
[0048] 1) Preparation of Standards
[0049] Collect goat milk and cow milk control substances, freeze-dry, and prepare goat milk containing 5 kinds of cow milk concentrations such as 100%, 50%, 25%, 10% and 1% (W / W) as standard substances for control, Total DNA was extracted, aliquoted and stored at -20°C for standard curve.
[0050] 2) Samples to be inspected
[0051] Commercially available goat milk powder was used as the test sample, and DNA was extracted with reference to Blood & Tissue Handbook.
[0052] 3) Primer and probe design and synthesis
[0053] The specific primers and probes of the internal control gene were designed for the homology sequence of 12SrRNA of cattle and sheep, and the primers and probes of specific genes of cattle were designed for the partial nucleotide sequence (SEQ ID NO.7) of 12SrRNA of cattle. Needle.
[0054] 12SrRNA is located on the mitochondria, which have the characteristics of high temper...
Embodiment 2
[0077] Embodiment 2: the analysis of inspection result
[0078] 1) Determination of standard curve and detection limit
[0079] Collect goat milk and cow milk control substances, freeze-dry, prepare goat milk containing 5 cow milk concentrations of 100%, 50%, 25%, 10%, 1% (W / W) and other control substances, and extract total DNA , taking the logarithmic values of the five standard concentrations as the horizontal axis, and the △Ct obtained by subtracting the average Ct value of the internal control gene Ct value from the average Ct value of the bovine-specific gene of each concentration standard product as the vertical axis, each concentration standard The product needs to be tested three times, and the difference between the Ct values of the parallel samples should be <0.5. Outliers should be discarded to ensure that there are representative data for each concentration before being included in the analysis and calculation.
[0080] The obtained formula model is: y=ax 2+...
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