Series: Brief measurement
What distinguishes the taste of Chinese soup?
Powdered Chinese soup stock is essential when making Chinese dishes. It is so convenient and easy to use♪ What is the secret of its taste?
We checked the type of meat extract using a mobile PCR device.
Suspension of 0.1 g of Chinese soup (made by company A) and 0.9 mL of purified water
Stirred using a vortex mixer.
1 μL was taken from near the center and was used as a sample.
|μL||Remarks (final concentration)|
|Enzyme: KAPA 3G Plant (2.5 U/μL)||0.8||2U/reaction|
|Primer F: beef/pork/chicken (100 μM each)||0.06/0.06/0.1||400/400/600nM|
|Primer R: beef/pork/chicken (100 μM each)||0.13/0.14/0.1||800/900/600nM|
|Probe: beef/pork/chicken (100 μM each)||0.03 each||200nM each|
|PCR grade water||4.7|
Reaction sample total 16μL
All indicators increased, indicating beef, pork and chicken are all included in the ingredients.
Real-time PCR amplification curves observed in the measurement are shown in the figure below.
Measurement was performed using a multiplex of beef, pork and chicken.
*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-BHQ1-3’ (25mer)||72.9|
|Pig-Primer F 5′-CTTGCAAATCCTAACAGGCCTG-3′ (22mer)||66.2|
|Pig-Primer R 5′-CGTTTGCATGTAGATAGCGAATAAC-3′ (25mer)||65.0|
|Pig-probe 5’ROX-AACAGCTTTCTCATCAGTTACACACAT-BHQ2-3′ (27mer)||70.2|
|Chicken-Primer F 5′-TCTGGGCTTAACTCTCATACTCACC-3′ (25mer)||65.6|
|Chicken-Primer R 5′-GGTTACTAGTGGGTTTGCTGGG-3′ (22mer)||65.6|
|Chicken-probe 5’Cy5-CATTCCTAACACTAGCCCTATTCTCC-3’BHQ3 (26mer)||70.3|
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