Series: Ernestly measurement
Dairy product residual volume in water discharged from the washing process of dairy production lines
These days, a single line is used for producing various products. Lines are cleaned after manufacturing a product and then used to manufacture a different product.
Some people may wonder if the line is actually clean. This is especially worrisome when dealing with allergens.
We checked the actual condition using a mobile PCR device.
[First step: Creation of a calibration curve]
Only dilution to achieve the following concentrations
106,104,103,102,101 pL/reaction as well as negative control
|μL||Remarks (final concentration)|
|Enzyme: KAPA 3G Plant (2.5 U/μL)||0.6||1.5U/reaction|
|F-Primer: Bovine (100 μM)||0.06||400nM|
|R-Primer: Bovine (100 μM)||0.13||800nM|
|Probe: Bovine (100 μM)||0.03||200nM|
|PCR grade water||5.4|
Reaction sample total 16μL
|Unit: pl/rxn||Ct value|
[Second step: Measurement of residual milk volume]
Water discharged from the cleaning process for dairy production lines
|Template||Ct||Residual milk volume
|Negative control||Not detected|
*1 Primer/probe sequence
|Cow-Primer F 5′-CCCGATTCTTCGCTTTCCAT-3′ (20mer)||66.8|
|Cow-Primer R 5’-CTACGTCTGAGGAAATTCCTGTTG-3′ (24mer)||65.4|
|Cow-probe 5’FAM-CATCATAGCAATTGCCATAGTCCAC-3’BHQ1 (25mer)||72.9|
Made by Nihon Gene Research Laboratories Inc.
Tm values are calculated using the nearest neighbor method. However, the calculation method of Nihon Gene Research Laboratories was used for the probes.
Reference paper:Tanabe et al., A Real-Time Quantitative PCR Detection Method for Pork, Chicken, Beef, Mutton, Horseflesh in Foods.
Biosci. Biotechnol. Biochem., 2007, 71(12),3131-3135