Addition of dNTPs can improve the detection sensitivity of catalytic hairpin assembly
Mingyuan ZouMeiling ZhouShuo MaChen ZhangXiao FengHuina WuGulinaizhaer AbudushalamuYuming YaoYaya ChenShijie CaiXiaobo FanFengfeng ZhaoGuoqiu Wu
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Abstract:
Ever since the catalytic hairpin assembly (CHA) circuit has been highlighted as a powerful nucleic acid detection tool, nucleic acid detection methods based on CHA have been widely studied. However, the detection sensitivity of CHA-based methods is insufficient. The relatively high background signals resulting from the spontaneous reaction between the two hairpin probes is one of the major reasons for limiting the sensitivity of CHA. In this study, we established that the addition of deoxynucleotide triphosphates (dNTPs) to the reaction system can significantly reduce the background leakage of CHA. The dNTPs-CHA, coupled with a fluorescence lateral flow assay strip, is used for the rapid and highly sensitive detection of miRNA. It is capable of reliably detecting miRNA in serum samples down to a limit of 100 aM, which is an improvement in the lower detection limit by nearly five orders of magnitude compared to that of the pure CHA.Keywords:
Limiting
The effects of four different temperature such as T1(18℃ day/12℃ night),T2(24℃ day/18℃ night),T3(30℃ day/24℃ night),T4(36℃ day/30℃ night) treatments on soluble protein and nucleic acid were studied in Chinese Kale.The results showed that the content of soluble protein was the highest in 36℃ day/30℃ night treatment and the lowest in 30℃ day/24℃ night treatment.The contents of total nucleic acid,DNA and RNA in the stem apex decreased with prolongation of temperature.with prolongation of temperature treatment was prolonged,the contents of total nucleic acid and RNA in all treatments rose gradually after decreasing except for 36℃ day/30℃ night treatment which decreased continuously.The content of DNA showed a changing trend such as decreasing-increasing-decreasing except for 36℃ day/30℃ treatment which increased after decreasing.Low temperature could accelerated the synthesis of nucleic acid,maintained the stability of the nucleic acid,but high temperature inhibited the synthesis of the nucleic acid,DNA and RNA,and subsequently led to the decreasing of nucleic acid contents.
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This chapter contains sections titled: Introduction Detection of nucleic acid targets by nucleic acid probes Functional nucleic acids Fluorescent nucleic acid sensors Colorimetric nucleic acid sensors Electrochemical nucleic acid sensors Piezoelectric nucleic acid sensors Conclusion and perspectives References
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Summary. Labelled sodium phosphate is administered to rats and after the lapse of 2 hours the specific activity of the ribo‐nucleic acid phosphorus and that of the desoxyribo‐nucleic acid phosphorus determined. In the liver the specific activity of the ribo‐nucleic acid P is found to be 33 times larger than the specific activity of the desoxyribo‐nucleic acid P. In the course of 2 hours about 0.1 and 3.3 per cent respectively of these compounds were found to be renewed. In the intestine and in the spleen in which the specific activity of the desoxyribo‐nucleic acid is found to be about 20 times larger than the corresponding value in the liver, the specific activity of the ribo‐nucleic acid phosphorus is only 2 to 3 times larger than the corresponding value of the desoxyribo‐nucleic acid phosphorus. The ribo‐and the desoxyribo‐nucleic acid phosphorus extracted from the total rat have a very similar specific activity to the corresponding phosphorus extracted from the intestine. In the total rat the difference in the rate of renewal of the two types of nucleic acid is not very pronounced. In a rat weighing 200 g approximately about 2 mg desoxyribo‐phosphorus and 3 mg ribo‐nucleic acid phosphorus are turned over in the course of 2 hours.
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Abstract The noncovalent interactions of night blue (NB) with several nucleic acids in buffer medium of Britton‐Robinson at pH 4.1 have been studied by spectroscopic methods. It is shown that the binding of NB with nucleic acids involves the J ‐aggregation of NB molecules on the surface of nucleic acids. The aggregation was encouraged by polyanions nucleic acids, in which nucleic acids served for acting templates. In this connection, a new method of nucleic acids with sensitivity at nanogram level is proposed based on the measurement of enhanced resonance light scattering (RLS). The linear range of ctDNA, fsDNA and yRNA is 0.01—2.5, 0.03—2.5 and 0.04—1.0 μg/mL, respectively, and the corresponding detection limits (3s̀) are 9.4, 7.3 and 5.7 ng/mL at 2.5 × 10– 5 mol/L of NB. Synthetic and real samples were analyzed with satisfactory results.
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CONTENTS I. Introduction 144 II. Structure and properties of nucleic acids 144 III. Effect of metal ions on the properties of nucleic acids 146 IV. Compounds of nucleic acids with metals 147 V. Structure of complexes of metals with nucleic acids 148 VI. Stability of complexes of metals with DNA 150
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The noncovalent interactions of night blue (NB) with several nucleic acids in buffer medium of Britton-Robinson at pH 4.1 have been studied by spectroscopic methods.It is shown that the binding of NB with nucleic acids involves the J-aggregation of NB molecules on the surface of nucleic acids.The aggregation was encouraged by polyanions nucleic acids,in which nucleic acids served for acting templates,In this connection,a new method of nucleic acids with sensitivity at nanogram level is proposed based on the measurement of enhanced resonance light scattering (RLS).The linear range of ctDNA,fsDNA and yRNA is 0.01-2.5,0.03-2.5 and 0.04-1.0 μg/mL,respectively,and the corresponding detection limits(3σ）are 9.4,7.3 and 5.7ng/mL at 2.5×10^05mol/L of NB.Synthetic and real samples were analyzed with satisfactory results.
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