2% (w/v) peqGOLD Universal Agarose (Peqlab) was dissolved in 1X TAE Buffer (Applichem) and 3 μL/100 mL Midori Green Advance (Biozym Scientific) were added. Products from RT-qPCR were mixed with Gel Loading Dye Blue (BioLabs). 100 bp Ladder (New England Biolabs) was used as reference. Gel electrophoresis was performed with 130 V for 40–120 min using the power supply unit peqPOWER E250 (Peqlab). DNA fragments were detected using FUSION-FX7 Spectra (Vilber).
Tae buffer
Manufactured by AppliChem
Sourced in Germany
TAE Buffer is a common buffer solution used in molecular biology laboratories. It is primarily utilized in agarose gel electrophoresis to separate and analyze DNA, RNA, and other nucleic acid molecules. The buffer solution maintains the appropriate pH and ionic conditions for the electrophoretic separation process.
Automatically generated - may contain errors
Lab products found in correlation
100 bp ladder, by New England Biolabs (1 mentions)
Peqgold universal agarose, by Avantor (1 mentions)
Fusion fx7 spectra, by Vilber (1 mentions)
Midori green advance, by Biozym (1 mentions)
Generuler low range dna ladder, by Thermo Fisher Scientific (1 mentions)
Tritrack dna loading dye, by Thermo Fisher Scientific (1 mentions)
Dna loading dye, by Thermo Fisher Scientific (1 mentions)
Peqgreen, by Avantor (1 mentions)
Loading dye, by Thermo Fisher Scientific (1 mentions)
Agarose, by AppliChem (1 mentions)
3 protocols using tae buffer
1
Agarose Gel Electrophoresis of RT-qPCR Products
2
Gel Electrophoresis for PCR Amplification
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Following the PCR, samples were tested for amplification success by gel electrophoresis using a 2% agarose gel (Sigma-Aldrich; A6877) in TAE buffer (AppliChem GmbH; A1691) at 4 °C. With a volume of 100 ml, the gel contained 4 µL of the dye peqGreen (VWR International GmbH; 37–500). For the loading of the agarose gel, 5 µL of the unpurified PCR product was mixed with 1 µL of 6× DNA Loading Dye (Thermo Fisher Scientific Inc.; R0611). The molecular weight marker used was composed of 1 µL Gene Ruler Low Range DNA Ladder (Thermo Fisher Scientific Inc.; SM1191), 1 µL 6 × TriTrack DNA Loading Dye (Thermo Fisher Scientific Inc.; R1161) and 4 µL nuclease-free water. Electrophoresis ran at a constant voltage of 100 V for 90–120 min (depending on the volume of the gel). The gel electrophoresis results were documented using the Quantum CX5 gel documentation system from Vilber Lourmat Deutschland GmbH (Eberhardzell, Germany) and the related BioVision software.
3
Gel Electrophoresis Visualization of PCR Products
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The result of the PCR was visualized by gel electrophoresis using a 2% agarose gel. 100 mL TAE-Buffer (AppliChem GmbH, Darmstadt, Germany) and 2 g agarose (AppliChem GmbH, Darmstadt, Germany) were taken, as well as 10 µL of the fluorescent dye GelRed (GeneON, Ludwigshafen, Germany). The loaded samples were composed of 5 µL of PCR product added to 1 µL loading dye (Thermo Fisher Scientific, Waltham, MA, USA). An electric field (90 mA, 90 V, 8 W) was applied for 2 h.
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