Total RNAs were isolated from HEK293 cells, stable HEK293 cells expressing eIF1Amutants (V55A, K56A, or K67A), and developing zebrafish with Trizol (Invitrogen). 25μg total RNAs were loaded into each well of TBE precast gel (Bio-Rad). Northern blot experiments were performed with semi-dry transfer cell and 5′-32P labeled probes. The mature miR-451 and miR-144 specific probes sequences are: 5′-AACUCAGUAAUGGUAACGGUUU-3′ for miR-451; 5′-AGUACAUCAUCUAUACU GUA-3′ for miR-144. U6 probe: 5′-AAAAAUAUGGAACGC UUCACGAAUUUG-3′ for U6 (probes were ordered from IDT).
Tbe precast gel
Manufactured by Bio-Rad
The TBE precast gel is a ready-to-use agarose gel designed for electrophoresis applications. It is pre-cast and pre-stained, providing a convenient solution for DNA and RNA separation and analysis.
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Lab products found in correlation
3 protocols using tbe precast gel
1
Northern Blot Analysis of miRNA Expression
2
Northern Blot Analysis of miRNA Expression
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Total RNAs were isolated from HEK293 cells, stable HEK293 cells expressing eIF1Amutants (V55A, K56A, or K67A), and developing zebrafish with Trizol (Invitrogen). 25μg total RNAs were loaded into each well of TBE precast gel (Bio-Rad). Northern blot experiments were performed with semi-dry transfer cell and 5′-32P labeled probes. The mature miR-451 and miR-144 specific probes sequences are: 5′-AACUCAGUAAUGGUAACGGUUU-3′ for miR-451; 5′-AGUACAUCAUCUAUACU GUA-3′ for miR-144. U6 probe: 5′-AAAAAUAUGGAACGC UUCACGAAUUUG-3′ for U6 (probes were ordered from IDT).
3
Electrophoresis and Autoradiography Protocol
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12 well TBE precast gel (BioRad) was pre-run at 150V for 20 minutes. The gel was loaded and run at 150V for 20–25 minutes, then gel was removed from the cassette, suspended between 2 sheets of cellophane, and dried. Gels were exposed to Biomax MR film (Carestream) in lead-lined cassettes at -80° and developed after 1–7 days.
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