Data for: Rapid detection of milk vetch dwarf virus by loop-mediated isothermal amplification(LAMP)

Published: 08-09-2018| Version 1 | DOI: 10.17632/7t4hzsvyr5.1
Contributors:
jun zhang,
Ying Li,
Tianbo Liu,
Zhicheng Zhou,
Shengping Wang,
Jinguang Yang,
Zhixin Xiao,
Jing Zhang,
Wei Li,
Songbai Zhang,
Fenglong Wang

Description

Figure captions Fig. 1. Optimization of the LAMP assay. F1a: The effect of the MgCl2 concentrations on the LAMP reaction at 63°C. Lanes 1–8 correspond to the 2, 3, 4, 5, 6, 7, 8 and 9 mM MgSO4 concentrations. F1b: The effect of the dNTPs on the LAMP reaction using 8 mM MgSO4. Lanes 1–7 correspond to the 0.8, 1.0, 1.2, 1.4, 1.6, 1.8 and 2.0 mM dNTP concentrations. F1c: The effect of temperature on the LAMP reaction using 8 mM MgSO4 and 1.6 mM dNTPs. Lanes 1–6 corresponsd to the temperatures of 60°C, 61°C, 62°C, 63°C, 64°C and 65°C. F1d: The effect of reaction time on the LAMP assay using 8 mM MgSO4 and 1.6 mM dNTPs at 65°C. Lanes 1–6 correspond to 15, 30, 45, 60, 75 and 90 min. M: DNA marker DL2000. Fig. 2. The effect of the ratio of the FIP and BIP primers to the F3 and B3 primers on the LAMP assay using 8 mM MgSO4 and 1.6 mM dNTPs at 65°C. Lanes 1–11 correspond to 1:1-11:1. M: DNA marker DL2000. Fig. 3. Test of the specificity of LAMP. Lane M, DNA marker DL2000; lane 1, healthy tobacco leaves; lane 2, MDV-infected tobacco leaves; lane 3, TYLCV-infected tobacco leaves; lane 4, TbCSV-infected tobacco leaves; lane 5, TbLCV-infected tobacco leaves; and lane 6, positive control. Fig. 4. Comparison of the relative sensitivities of LAMP and PCR for MDV detection. F4a: lane 1, distilled water; lanes 2-8, DNA concentrations of 100, 10, 1, 0.1, 0.01, 0.001 and 0.0001 ng. F4b: lane 1, distilled water; lanes 2-8, DNA concentrations of 100, 10, 1, 0.1, 0.01, 0.001 and 0.0001 ng. Lane 9, positive control. M: DNA marker DL2000. Fig. 5. The detection of natural viral infection in selected field-grown tobacco samples using LAMP. F5a: lane 1, distilled water; lanes 2-12 tobacco samples showing MDV or virus-like symptoms; lane 13: positive control. F5b: Visual inspection of the MDV LAMP products. All products in the reaction tubes were stained with SYBR Green I. A positive reaction appeared green and a negative reaction appeared orange in daylight. F5c: A positive reaction was indicated by green fluorescence and a negative reaction was indicated by fluorescence under a UV lamp. The number of F5b and F5c is consistent with F5a. M: DNA marker DL2000.

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