Arabidopsis Genomic DNA Southern Blotting

Southern blotting analysis was carried out as described (Cheng et al., 2015 (link)). Briefly, 20 μg Arabidopsis genomic DNA was digested with EcoRI. The DNA was resolved on a 0.8% agarose gel and transferred onto an Amersham Hybond^TM-N⁺ nylon membrane (GE Healthcare, USA); the blot was cross-linked under 0.12 J/cm² UV irradiation. Then the blot was hybridized at 42°C with Digoxigenin-labeled probe for 12 h. After two stringent washes at 68°C, the signal was detected using DIG Nucleic Acid Detection Kit (Roche, USA).

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Cheng H., Chen X., Zhu J, & Huang H. (2016). Overexpression of a Hevea brasiliensis ErbB-3 Binding protein 1 Gene Increases Drought Tolerance and Organ Size in Arabidopsis. Frontiers in Plant Science, 7, 1703.

Publication 2016

Amersham HybondTM-N+ nylon membrane DIG Nucleic Acid Detection Kit

Agarose Arabidopsis Digoxigenin Ecori Genomic Membrane Nucleic acid Nylon Southern blotting analysis Uv irradiation

Corresponding Organization : Rubber Research Institute

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Variable analysis

independent variables

Digoxigenin-labeled probe

dependent variables

Signal detected using DIG Nucleic Acid Detection Kit

control variables

20 μg Arabidopsis genomic DNA
DNA digested with EcoRI
DNA resolved on 0.8% agarose gel
DNA transferred onto Amersham Hybond™-N+ nylon membrane
Blot cross-linked under 0.12 J/cm^2 UV irradiation
Blot hybridized at 42°C for 12 h
Two stringent washes at 68°C

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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