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What distinguishes the taste of Chinese soup?

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.

Sample

Suspension of 0.1 g of Chinese soup (made by company A) and 0.9 mL of purified water

Pretreatment

Stirred using a vortex mixer.
1 μL was taken from near the center and was used as a sample.

Reaction composition
μL Remarks (final concentration)
Enzyme: KAPA 3G Plant (2.5 U/μL) 0.8 2U/reaction
Buffer (x2) 8 x1
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
MgCl2(25mM) 0.8 1.25mM
Sample 1
PCR grade water 4.7

Reaction sample total 16μL

Program
Process Temperature Time
Initial denaturation 95℃ 15sec
Annealing/extension 64℃ 20sec 45cycle
Denaturation 95℃ 4sec

 

Results

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
Tm
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