Primer probe group and kit for human leukotriene receptor CysLTR2 mRNA RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection
A primer-probe, leukotriene technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of small detection range, problems with accuracy, low sensitivity, etc.
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Embodiment 1
[0036] 1. The reagents and equipment involved are as follows:
[0037] 1.1 Reagents
[0038] 1.1.1 Whole blood total RNA kit (Hangzhou Xinjing Biological Reagent Development Co., Ltd., catalog number: 5201050)
[0039] 1.1.2 HiScribe T7 High Yield RNA Synthesis Kit (New England Biolabs, Cat. No.: E2050S)
[0040] 1.2 Main instruments
[0041] 1.2.1 Applied Biosystems TM 7300 fluorescent quantitative PCR instrument: ThermoFisher, USA
[0042] 1.2.2-80℃ low-temperature refrigerator: ThermoFisher, USA
[0043] 1.2.3 High-speed and low-temperature desktop centrifuge: Eppendorf, Germany
[0044] 1.2.4 Qubit 3 Fluorometer: ThermoFisher, USA
[0045] 2. Method
[0046] 2.1 Primer and probe design
[0047] According to the sequences of CysLTR2 and GAPDH, use Primer 6.0 software to design fluorescent quantitative primers and probes. After a series of effect verification, the primer pairs CysLTR2-F, CysLTR2-R, GAPDH-F, GAPDH-R and probes of CysLTR2 and GAPDH were obtained. E-Pr...
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