Native polyacrylamide gel electrophoresis (PAGE) was performed in a temperature-controlled vertical electrophoretic apparatus (Z375039-1EA; Sigma-Aldrich, San Francisco, CA). Gel concentration was 10% (19:1 monomer to bis ratio, AppliChem, Darmstadt). Approximately 2 μg of sample was loaded onto 14 × 16 × 0.1 cm gels. Electrophoreses were performed at 37 °C for 3 h at 126 V (~8 V cm−1). DNA oligomers were visualised with Stains-all immediately after the electrophoresis, and the electrophoretic record was photographed on a white pad with a Nikon D3100 camera. Additional staining with silver was performed consequently. The modified Britton-Robinson buffer was used in all electrophoretic separations; pH was adjusted by TRIS to a final value of 7.6.
Z375039 1ea
Manufactured by Merck Group
Sourced in United States
The Z375039-1EA is a laboratory equipment product. It serves as a core function in laboratory settings, but a detailed description without interpretation or extrapolation is not available.
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Lab products found in correlation
5 protocols using z375039 1ea
1
Native PAGE Analysis of Biomolecules
2
Non-denaturing PAGE Analysis of DNA
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Samples consisting of 0.3 μl of 1 mM stock solutions were separated using nondenaturing PAGE in a temperature-controlled electrophoretic apparatus (Z375039-1EA; Sigma-Aldrich, San Francisco, CA) on 15% acrylamide (19 : 1 acrylamide/bisacrylamide) gels. DNA was loaded onto 13 × 16 × 0.1 cm gels. Electrophoresis was run at 10°C for 4 hours at 125 V (~8 V·cm−1). Each gel was stained with StainsAll (Sigma-Aldrich). The gel was also stained using the silver staining procedure in order to improve the sensitivity of the DNA visualization [44 (link)].
3
Native PAGE Analysis of DNA Oligomers
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Native polyacrylamide gel electrophoresis (PAGE) was performed in a temperature controlled vertical electrophoretic apparatus (Z375039-1EA, Sigma-Aldrich, San Francisco, CA). Gel concentration was 12% (19 : 1 monomer to bis ratio, Applichem, Darmstadt). Approximately two micrograms of DNA was loaded onto 14 × 16 × 0.1 cm gels. Prior to loading, each DNA sample was heated to 95°C for 5 min in an appropriate buffer and cooled to room temperature. Electrophoreses were performed at 20°C for 4 hours at 120 V (~8 V·cm−1). DNA oligomers were visualized with Stains-All immediately after electrophoresis, and the electrophoretic record was photographed on a white pad with a Nikon D3100 camera. The gel was also later stained by silver staining procedure in order to improve the sensitivity of the DNA visualization [15 (link), 35 (link)].
4
Separation and Visualization of DNA Structures
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Samples consisting of 0.3 µL of 1mM stock solutions were separated using nondenaturing PAGE in a temperature-controlled electrophoretic apparatus (Z375039-1EA; Sigma-Aldrich, San Francisco, CA, USA) on 12% acrylamide (19: 1 acrylamide/bisacrylamide) gels. DNA was loaded onto 13 by 16 by 0.1 cm gels. Electrophoresis was run at 10 °C for 2 h at 125V (~8 V⋅cm−1). Each gel was stained with StainsAll (Sigma-Aldrich). All electrophoretic measurements were performed in a mBR buffer at pH 7.0. Temperature gradient gel electrophoresis (TGGE) equipment was used according to a method which has been described previously [44 (link),45 (link)]. The gel concentration was 12%. Electrophoreses were run perpendicularly to the temperature-gradient (20–80 °C) for 3 h at 160 V (~8 V·cm−1). Approximately 12 μg of DNA was loaded into the electrophoretic well. DNA oligomers were visualized with Stains-all after the electrophoresis. Ligand gradient gel electrophoresis (LGGE) is similar to denaturing gradient gel electrophoresis (DGGE), but in place of a denaturing agent, a concentration gradient of ligand (0–260 µM) is applied perpendicularly to the movement of the DNA sample. The same apparatus used for standard PAGE analyses was used in this assay. The technique was developed and applied in our laboratory to monitor the folding and multimerization effect of the ligands on G4 structures.
5
Nondenaturing PAGE and TGGE Separation
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Samples consisting of ~0.3 µL of 1 mM stock solutions were separated using nondenaturing PAGE in a temperature-controlled electrophoretic apparatus (Z375039-1EA; Sigma-Aldrich, San Francisco, CA, USA) on 12% acrylamide (19: 1 acrylamide/bisacrylamide) gels. Electrophoresis was run at 15° and 60°C for 2 h at 125 V (~8 V·cm−1). All electrophoretic measurements were performed in the mBR buffer at pH 7.0. Temperature gradient gel electrophoresis (TGGE) equipment was used according to the previously described method [42 (link),47 (link)]. As in the previous study, the gel concentration was 12%. Electrophoreses were run perpendicularly to the temperature gradient (15–80 °C) for 3 h at 160 V (~8 V·cm−1). Approximately 12 μg of DNA was loaded into the electrophoretic well. Each gel was stained with StainsAll (Sigma-Aldrich).
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