Application one,
Hou Xiaofeng from the Department of Biochemistry of PLA General Hospital conducted methodological evaluation and performance verification on the detection of lactate dehydrogenase isoenzyme 1 (LDH1) by guanidine thiocyanate inhibition method. Methods: The imprecision, linear analysis, recovery experiment and normal reference range of LDH1 detection by guanidine thiocyanate inhibition method were evaluated. Results: The intra-assay imprecision of high, medium and low value samples were 4.25%, 1.57% and 0.97%, respectively, and the inter-assay imprecision were 4.57%, 2.55% and 1.20%, respectively; the dilution robustness of the linear analysis was good, R2 =0.998; the average recovery rate was 101.56%; there was no significant difference in LDH1 level between genders; the normal reference range was 33.11±5.76U/L. Conclusion: The guanidinium thiocyanate inhibition method for detecting LDH1 is reliable, has good performance, meets the laboratory requirements, and can be widely used in clinical practice.
Application two,
CN201110146840.5 discloses a method for preparing total RNA without genomic DNA, using sodium dodecyl sulfate as a lysis reagent, and using potassium acetate and sodium dodecyl sulfate to react to generate potassium dodecyl sulfate (PDS) after lysing cells Precipitates, co-precipitates proteins as well as most of the genomic DNA. Then, utilizing the property that nucleic acid can be adsorbed by the phenol extract when guanidinium thiocyanate exists, and the adsorbed nucleic acid can be dissociated by a certain concentration of potassium acetate according to the molecular weight, a certain concentration of guanidinium thiocyanate and acidic acetic acid are added to the nucleic acid. Potassium, total RNA and residual genomic DNA were isolated by phenol extraction. Finally, total RNA samples were obtained by ethanol precipitation. Compared with the prior art, this method has the least residual genomic DNA in the obtained RNA sample, which is 103 copies per microgram (about 4 picograms).
Application three.
CN201711222569.2 provides a lysate for extracting nucleic acid from exfoliated cells in human feces, including: 1mol/L~8mol/L guanidinium salt, 1mM~20mM metal ion chelating agent and 100mM-Tris-HCl buffer solution with pH7.5~8.0, Wherein, the guanidine salt is selected from at least one of guanidine hydrochloride, guanidine isothiocyanate and guanidine thiocyanate, and the metal ion chelating agent is selected from at least one of ethylenediaminetetraacetic acid and water-soluble salts of ethylenediaminetetraacetic acid. kind. Preferably, it also includes 1% volume to 20% volume of surfactant, the surfactant is a non-ionic detergent, and the non-ionic detergent is selected from Tween-20, Tween-80, Brij-35, NP40 and At least one of Triton-100. Related preparation methods and applications are also provided. The lysate of the present invention can rapidly and efficiently lyse cells, fully release nucleic acid, meet the amount of subsequent nucleic acid detection, and is suitable for large-scale popularization and application.