Lab 09-GUS-DNA-PCR - Lab 9 Reporter Genes GUS The GUS gene...

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Lab 9 Reporter Genes GUS The GUS gene is used in transgenic studies to verify the transmission of the transformation vector into the target plant. It is informative because nothing in the plant tissue would stain blue; the blue stain can only mean the plant is transformed. GUS makes a protein that turns blue when mixed with X-gluc (5-Brom-4-chlor-3-indolyl-ß- Dglucuronide). Chimeras can be observed in the X-gluc stained tissue. X-gluc stain (prepared): 2.5ml 0.2M Na 2 HPO 4 2.4ml D.I. water 100µl 0.5M Na 2 EDTA 5.0mg X-gluc 5ml 1. Put 30-40µL fresh X-gluc stain into the well of a microtiter plate. 2. Select a shoot from your Tobacco Transformation Experiment. Cut a thin section of leaf or stem and place it into an individual well - keep track of where sections come from. 3. Stain sections for 4-5 hr. at 37 ° in the incubator. 4. After staining, clear or fix the tissue by adding 50µL of 95% ethanol:glacial acetic acid (3:1 v/v). Wait a half hour before scoring for GUS. Perform step under
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Lab 09-GUS-DNA-PCR - Lab 9 Reporter Genes GUS The GUS gene...

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