Kit for detecting chronic sinusitis with subtype nasal polyp as well as application of CLC gene as biomarker
A chronic sinusitis and kit technology, applied in the field of biomedicine, can solve problems such as difficult batch operation, time-consuming, patient fear, etc., to achieve the effect of saving labor costs, saving medical costs, and solving human errors
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Embodiment 1
[0068] A kit for detecting subtypes of chronic sinusitis with nasal polyps, comprising the following reagents:
[0069] Reagents for extracting RNA from nasal polyp tissue: 20mL RNA extraction solution Trizol or RNAiso Blood or RNAiso Plus or others containing phenol, guanidine isothiocyanate, 8-hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol, etc. Substances that can rapidly disrupt cells and inhibit nucleases released from cells; 2mL of chloroform; 20mL of isopropanol; 40mL of 65-90% ethanol; 5mL of RNase- and DNase-free water;
[0070] Reagents for reverse transcription of extracted RNA into cDNA: 40 μL of reverse transcription mixture (containing enzymes required for reverse transcription, RNase inhibitors, random 6-nucleotide primers, polythymine, T repeat oligonucleotides , deoxyribonucleotide triphosphate mixture, buffer, etc.), 160 μL of RNase- and DNase-free water; the RNase- and DNase-free water is used to make up the system, dissolve, and dilute RNA; ...
Embodiment 2
[0073] A kit for detecting subtypes of chronic sinusitis with nasal polyps, comprising the following reagents:
[0074] Reagents for extracting RNA from nasal polyp tissue: 1mL RNA extraction solution Trizol or RNAiso Blood or RNAisoPlus or other substances containing phenol, guanidine isothiocyanate, 8-hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol, etc. Substances that can rapidly disrupt cells and inhibit nucleases released from cells; 0.2 mL of chloroform; 0.2 mL of isopropanol; 0.2 mL of 65-90% ethanol; 0.05 mL of RNase- and DNase-free water;
[0075] Reagents for reverse transcription of extracted RNA into cDNA: 2 μL of reverse transcription mixture (containing enzymes required for reverse transcription, RNase inhibitors, random 6-nucleotide primers, polythymine, T repeat oligonucleotides , deoxyribonucleotide triphosphate mixture, buffer, etc.), 7 μL of RNase- and DNase-free water; the RNase- and DNase-free water is used to make up the system, dissolve, a...
Embodiment 3
[0078] A kit for detecting subtypes of chronic sinusitis with nasal polyps, comprising the following reagents:
[0079] Reagents for extracting RNA from nasal polyp tissue: 0.1mL RNA extraction solution Trizol or RNAiso Blood or RNAiso Plus or other containing phenol, guanidine isothiocyanate, 8-hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol 0.05mL of chloroform; 0.015mL of isopropanol; 0.0075mL of 65-90% ethanol; 0.01mL of RNase- and DNase-free water ;
[0080] Reagents for reverse transcription of total RNA into cDNA: 1 μL of reverse transcription mixture (containing enzymes required for reverse transcription, RNase inhibitors, random 6-nucleotide primers, polythymine, T repeat oligonucleotides, Deoxyribonucleotide triphosphate mixture, buffer, etc.), 0-10 μL of RNase- and DNase-free water; the RNase- and DNase-free water is used to make up the system, dissolve, and dilute RNA;
[0081] Reagents for real-time fluorescent quantitative PCR reaction of CLC gene...
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