Specific primers, probes, kits and methods for detecting barley components
A kit and specific technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problem of inability to distinguish barley from other species, achieve short time, accurate results, and identification The results are accurate
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Embodiment 2
[0061] Example 2 Probe
[0062] This embodiment provides a probe for detecting barley components. The probe designed for the ACPS gene of barley is as follows, as shown in SEQ ID NO: 3 in the sequence listing: the sequence of the probe is: 5'-TCGCGCCATTCAAGCCGTGATACGTC -3'.
Embodiment 3
[0063] Example 3 Common PCR Detection Kit
[0064] This embodiment provides a test kit for detecting barley components, each of which includes a separately packaged primer solution part and a PCR reaction solution part; the primer solution part contains the The specific primer; the PCR reaction solution part contains amplification buffer, dNTPs, Taq DNA polymerase and sterile ultrapure water for PCR amplification, and the formula of each kit is:
[0065] Upstream primer, 10μM, 0.5-1.5μL;
[0066] Downstream primer, 10μM, 0.5-1.5μL;
[0067] Taq DNA polymerase, 5U / μL, 0.2~1.2μL;
[0068] 10×Taq DNA polymerase buffer (containing 10mmol / L Mg 2+ ), 2.5μL;
[0069] dNTPs, 2.5mM, 0.25-1μL; add sterile double distilled water to a total volume of 25μL.
Embodiment 4
[0070] Embodiment 4 real-time fluorescent PCR detection kit
[0071] This embodiment provides a test kit for detecting barley components, each of which includes a separately packaged primer solution part and a PCR reaction solution part; the primer solution part contains the The specific primers and the probe described in Example 2; the PCR reaction solution part contains 2 × TaqMan Universal PCR Master Mix containing dNTPs, Taq DNA polymerase and sterile ultra- Pure water, the formula of each said kit is:
[0072] 2×TaqMan Universal PCR Master Mix 12.5μL;
[0073] Upstream primer (10 μM) 1.0 μL;
[0074] Downstream primer (10 μM) 1.0 μL;
[0075] Probe (10 μM) 0.5 μL;
[0076] Add sterile double distilled water to a total volume of 25 μL.
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