Specific primers, probes, kits and methods for detecting sugarcane components
A kit and probe technology, applied in the field of probes, kits, and specific primers for detecting sugarcane components, can solve the problem of inability to distinguish sugarcane from other species, and achieve the effects of small molecular weight, strong specificity, and high sensitivity
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Embodiment 2
[0066] Example 2 Probe
[0067] This embodiment provides a probe for detecting sugarcane components. The probe designed for the ACPS gene of sugarcane is as follows, see SEQ ID NO: 3 in the sequence listing, the sequence of the probe is: 5'-CGATCGATGTTCTGGGTGCTGAGTGCC -3'.
Embodiment 3
[0068] Example 3 Common PCR Detection Kit
[0069] The present embodiment provides a kind of test kit for detecting sugarcane components, and each said kit includes a primer solution part and a PCR reaction solution part of independent packaging; 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:
[0070] Upstream primer (10 μM) 1.0 μL;
[0071] Downstream primer (10 μM) 1.0 μL;
[0072] Taq DNA polymerase, 5U / μL, 0.4μL;
[0073] dNTPs, 10mM, 0.5μL;
[0074] 10×Taq DNA polymerase buffer (containing 10mmol / L Mg 2+ ), 2.5μL;
[0075] Make up to 25 μL with sterile ultrapure water;
[0076] Add sterile double distilled water to a total volume of 25 μL.
Embodiment 4
[0077] Embodiment 4 real-time fluorescent PCR detection kit
[0078] The present embodiment provides a kind of test kit for detecting sugarcane components, and each said kit includes a primer solution part and a PCR reaction solution part of independent packaging; The specific primers and the probe described in Example 2; the PCR reaction solution part contains 2×TaqManUniversal PCR Master Mix of dNTPs, Taq DNA polymerase and sterile ultrapure water, the formula of each kit for:
[0079] 2×TaqMan Universal PCR Master Mix 12.5μL;
[0080] Upstream primer (10 μM) 1.0 μL;
[0081] Downstream primer (10 μM) 1.0 μL;
[0082] Probe (10 μM) 0.5 μL;
[0083] Add sterile double distilled water to a total volume of 25 μL.
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