The fungus was cultured for seven days onto potato dextrose broth medium. The mycelium was separated and ground and the total genomic DNA was isolated from the fungal mycelium according to [22 ]. The isolated DNA was resolved on 1% agarose gel (Sigma-Aldrich) prepared in 1 X TAE (Tris-acetate-ethylenediaminetetra acetic acid) buffer containing 0.5 µg ml−1 ethidium bromide (Sigma-Aldrich). ethidium bromide-stained gel was visualized using a UV-transilluminator (Vilber Lourmat, Collégien, France).
Ethidium bromide
Manufactured by Vilber
Sourced in France
Ethidium bromide is a nucleic acid stain used in molecular biology to visualize DNA and RNA in agarose gels. It binds to nucleic acids and emits fluorescence under ultraviolet light, allowing the detection of DNA or RNA bands in electrophoresis gels.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ethidium bromide
1
Fungal Genomic DNA Isolation and Visualization
2
RT-PCR Analysis of MUC5AC Gene Expression
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RNA was extracted using TRIZOL reagent (Qiagen) according to the manufacturer's protocol. The RNA concentration was quantified by Biophotometer (Eppendorf, Germany). cDNAs were synthesized from RNA in the presence of M-MuLV reverse transcriptase (No. 610600900021730 Merck–Mumbai). Specific primer sequence for MUC5AC was used to amplify gene transcripts. The reaction was run in a thermal cycler (Eppendorf, Germany). The PCR conditions were 5 min of initial denaturation at 95°C and 36 cycles consisting of 45 s of denaturation at 95°C followed by 1 min at 59°C annealing step and 1 min at 72°C elongation steps. The final 10 min incubation at 72°C assured a complete extension of the PCR products. The presence of amplified products were electrophoresed on 1% agarose gel with ethidium bromide and visualized in UV light (Vilber Lourmat, France). The primers (Eurofins scientific, Bangalore, India) used were listed in Table S1 and S2 .
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