Series: Brief measurement
Does milk have any cow genes?
Milk is nutritious and reasonable. There are a lot of types of milk products including low-fat milk, processed milk, and dairy products. Is the difference measurable?
Testing the difference is important for people with allergies.
Measuring milk is very interesting because milk is very familiar to our life. We first checked if milk has any cow genes using a mobile PCR device.
|μL||Remarks (final concentration)|
|Enzyme: KAPA 3G Plant (2.5 U/μL)||0.6||1.5U/reaction|
|F-Primer: cow (100 μM)||0.06||400nM|
|R-Primer: cow (100 μM)||0.13||800nM|
|Probe: cow (100 μM)||0.03||200nM|
|PCR grade water||5.4|
Reaction sample total 16μL
Cow genes were present in an amount that the sensor easily reacted even when it was diluted 100 fold.
This indicates that the presence of milk residue can be measured via PCR.
*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