RNA SNP Detection Method With Improved Specificity Based on Dual-competitive-padlock-probe

Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.

Study of beam divergence and thrust vector eccentricity characteristics of the Hall thruster based on dual Faraday probe array planes and its applications

The accurate knowledge of the thrust vector eccentricity and beam divergence characteristics of Hall thrusters are of significant engineering value for the beneficial integration and successful application of Hall thrusters on spacecraft.For the characteristics of the plume bipolar diffusion due to the annular discharge channel of the Hall thruster,a Gaussian-fitted method for thrust vector deviation angle and beam divergence of Hall thrusters based on dual Faraday probe array planes was proposed in respect of the Hall thruster beam characteristics.The results show that the ratios of the deviation between the maximum and minimum values of the beam divergence angle and the thrust vector eccentricity angle using a Gaussian fit to the optimized Faraday probe dual plane to the mean value are 1.4%and 11.5%,respectively.The optimized thrust vector eccentricity angle obtained has been substantially improved,by approximately 20%.The beam divergence angle calculated using a Gaussian fitting to the optimized Faraday probe dual plane is approximately identical to the non-optimized one.The beam divergence and thrust vector eccentricity angles for different anode mass flow rates were obtained by averaging the beam divergence and thrust vector eccentricity angles calculated by the dual-plane,Gaussian-fitted ion current density method for different cross-sections.The study not only allows for an immediate and effective tool for determining the design of thrust vector adjustment mechanisms of spacecraft with different power Hall thrusters but also for characterizing the 3D spatial distribution of the Hall thruster plume.

Plasma diagnosis of tetrahedral amorphous carbon films by filtered cathodic vacuum arc deposition

Filtered cathodic vacuum arc(FCVA)deposition is regarded as an important technique for the synthesis of tetrahedral amorphous carbon(ta-C)films due to its high ionization rate,high deposition rate and effective filtration of macroparticles.Probing the plasma characteristics of arc discharge contributes to understanding the deposition mechanism of ta-C films on a microscopic level.This work focuses on the plasma diagnosis of an FCVA discharge using a Langmuir dualprobe system with a discrete Fourier transform smoothing method.During the ta-C film deposition,the arc current of graphite cathodes and deposition pressure vary from 30 to 90 A and from 0.3 to 0.9 Pa,respectively.The plasma density increases with arc current but decreases with pressure.The carbon plasma density generated by the arc discharge is around the order of10^(10)cm^(-3).The electron temperature varies in the range of 2-3.5 eV.As the number of cathodic arc sources and the current of the focused magnetic coil increase,the plasma density increases.The ratio of the intensity of the D-Raman peak and G-Raman peak(I_(D)/I_(G))of the ta-C films increases with increasing plasma density,resulting in a decrease in film hardness.It is indicated that the mechanical properties of ta-C films depend not only on the ion energy but also on the carbon plasma density.