Northern Blot Analysis of RNA Transcripts

RNAs were isolated with RNA extract kit. Northern blot analysis was performed with northern blot kit (Ambion) as described (27 (link)). Briefly, the preparation of total RNAs (30 μg) was denatured in formaldehyde and then electrophoresed in a 1% agarose–formaldehyde gel. The RNAs were then transferred onto a Hybond-N+nylon membrane (Amersham) and hybridized with biotin-labeled DNA probes. Biotin Chromogenic Detection kit (Thermo Scientific) was used to develop the bound RNAs.

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Du W.W., Yang W., Liu E., Yang Z., Dhaliwal P, & Yang B.B. (2016). Foxo3 circular RNA retards cell cycle progression via forming ternary complexes with p21 and CDK2. Nucleic Acids Research, 44(6), 2846-2858.

Publication 2016

northern blot kit Hybond-N+nylon membrane Biotin Chromogenic Detection kit

Agarose Biotin Chromogenic Dna probes Formaldehyde Membrane Northern blot Northern blot analysis Nylon Rnas

Corresponding Organization :

Other organizations : Sunnybrook Health Science Centre, Health Sciences Centre, University of Toronto

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Variable analysis

independent variables

RNA extraction kit

dependent variables

RNA expression levels (as measured by northern blot analysis)

control variables

Amount of total RNA (30 μg) used for northern blot analysis
Agarose-formaldehyde gel used for electrophoresis
Hybond-N+ nylon membrane used for RNA transfer
Biotin-labeled DNA probes used for hybridization
Biotin Chromogenic Detection kit used for developing the bound RNAs

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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