The RT-PCR products were separated by electrophoresis in a 1.5% agarose gel in 0.5 × Tris–borate–ethylenediaminetetraacetic acid (TBE) buffer (SERVA, Heidelberg, Germany) stained with GelRed nucleic acid stain (Phenix Research Products, Candler, NC, USA). Each well was loaded with 5 μl of the PCR product and 1 μl of 6 × blue–orange DNA loading dye (Promega, Madison, USA). Samples were separated along with DNA ladder (Promega, Madison, USA) at 150 V for 30 min. The PCR products were visualized using a gel documentation system (EZ Gel Doc, Bio-Rad, France) and scoring was done based on the size of the PCR products.
Blue orange dna loading dye
Manufactured by Promega
Sourced in United States
Blue–orange DNA loading dye is a solution used to visualize DNA samples in gel electrophoresis. It contains blue and orange dyes that allow the DNA bands to be easily identified and tracked during the electrophoresis process.
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Lab products found in correlation
2 protocols using blue orange dna loading dye
1
Agarose Gel Electrophoresis of RT-PCR Products
2
Agarose Gel Electrophoresis of PCR Amplicons
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The PCR products were separated by electrophoresis on a 1.5% agarose gel (UltraPure Agarose; Invitrogen Life Technologies) in 1 × Tris–borate–ethylenediaminetetraacetic acid (TBE) stained with GelRed nucleic acid stain (Phenix Research Products, Candler, NC, USA). Each well was loaded with 9 μl of amplicons and 1.5 μl of 6 × blue–orange DNA loading dye (Promega, Madison, WI, USA). The gel was run at 200 V for 30 min and the PCR products were visualized under UV light using a gel imager camera (Red Personal Gel Imaging System; Alpha Innotech, USA). The size of positive PCR products was estimated to be around 600 bp by comparing the amplicons with a DNA ladder, as shown in Figure 2 .Figure 2 ![]()
Agarose gel electrophoresis of PCR showing the 200-bp DNA ladder (M), DNA samples (lanes 1–8), negative control (NC) and positive control (PC) for B19V DNA.
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