Phlegm resolving liquid for extracting free DNAs from cell-free supernatant of sputum specimen
A cell-free and sputum technology, applied in the field of sputum removal, can solve the problems of low cfDNA content in cell-free supernatant, affecting the accuracy of gene detection results, etc., and achieve the effects of abundant cfDNA content, easy availability, and widespread promotion.
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[0042] Example 1,
[0043] The formula of the phlegm solution is determined: 1) It is convenient for clinical and laboratory acquisition; 2) It must be an isotonic liquid to ensure that the sputum cytology is intact and does not affect the cytological pathological diagnosis; 3) Minimize the phlegm solution in the supernatant The salt ion content of the supernatant avoids affecting the extraction efficiency of the cfDNA kit extracted from the supernatant; 4) Dithiothreitol and β-mercaptoethanol are commonly used denaturants in experiments, but dithiothreitol is colorless and tasteless and more suitable for laboratories Use; 5) The literature reports that there is a big difference in the concentration of DTT to resolve phlegm [1]. Under different salt ion concentrations, the optimal concentration of dithiothreitol requires experimental proof.
[0044] In order to observe whether the concentration of dithiothreitol changes the phlegm-reducing effect, this experiment was conducted with...
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