High-throughput multi-LDR parting kit for detecting farmland ecological environment
An eco-environmental, high-throughput technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as difficult to achieve high-throughput detection
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[0109] (1) Amplification of the DNA of the tarantula and the DNA of the prey with universal primers
[0110] (1) Mix the standard plasmids of 16 species at equal concentrations, and the total concentration is 10 pg / μL;
[0111] (2) Universal primers
[0112] Upstream: 5′-GGTCAACAAATCATAAAGATATTGG-3′;
[0113] Downstream: 5′-TAAACTTCAGGGTGACCAAAAAAATCA-3′;
[0114] (3) Amplification system (10μl):
[0115] Upstream primer 0.5μM
[0116] Downstream primer 0.5μM
[0117] 10×Buffer 1μl
[0118] dNTP 0.2mM
[0119] Mg 2+ 2.0mM
[0120] Taq polymerase 0.1μl (0.5Unit)
[0121] Add water to 10μl;
[0122] (4) Amplification program:
[0123] 95℃15min
[0124] 94℃30s→50℃1min→72℃2min(35cycles)
[0125] 72℃7min→4℃forever
[0126] (2) Ligase detection reaction (LDR) between the DNA of the tarantula and the DNA of the prey
[0127] (1) Use the PCR product amplified by the universal primer as a template for LDR signal detection
[0128] (2) LDR system (10μl):
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