An experiment was performed to analyze the effects of heat shock in yeast called Saccharomyces cerevisiae, to assess the hypothesis that yeast extracts up-regulate gene expression during heat shock the same way bacteria cells up-regulate gene expression during heat shock. In bacteria cells, heat shock causes the bacteria to up-regulate gene expression and produce more proteins. Similarly, gene expression and protein production increases in yeast during heat shock. Therefore, the experiment was conducted to test the expression of a particular gene, called HSP 30, in yeast (Saccharomyces cerevisiae), grown in liquid culture. Yeast strains with GFP-tagged transgenes were used. The purpose of the GFP tag on the gene was to detect the proteins produced
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To obtain similar but more accurate results, a lower amount of protein extract should be loaded into the wells of the polyacrylamide gel. The same procedure will be followed, except there will be less protein extract in each of the wells (lanes 1-4). Instead of loading 10 ug of protein extract, only 5 ug will be loaded to each well. The hypothesis of this experiment would be the same that yeast extracts up-regulate gene expression during heat shock the same way bacteria cells up-regulate gene expression during heat shock. Adding a lower amount of protein extract will produce fewer bands, but will eliminate the cluster of bands seen in lanes 1 and 3 of the current experiment (Fig. 1). In lane 3 of figure 1, instead of having a cluster of bands around 64KD, there might only be 1 band present at 64KD. The experiment addresses a technical question because a small part of the procedure is getting changed in order to improve the results. The technical question the experiment proposes is: are their bands present when 5 ug of protein extract are added to each well? If the experiment supports the hypothesis, then the lane with HSP 30 gene at 42°C will have one band at approximately 64 KD rather than a cluster of bands. This means a clear and accurate conclusion can be drawn from the results. The presence of one band at around 64 KD will still conclude that the expression of the HSP 30 gene is greater at 42°C than 30°C. However, if a band is not present around 64KD, then the results would not support the hypothesis. Overall, the amount of protein extract added to each sample, before conducting gel electrophoresis, will pose a technical question that can improve the results and accurately conclude whether HSP 30 gene expression increases in heat
To obtain similar but more accurate results, a lower amount of protein extract should be loaded into the wells of the polyacrylamide gel. The same procedure will be followed, except there will be less protein extract in each of the wells (lanes 1-4). Instead of loading 10 ug of protein extract, only 5 ug will be loaded to each well. The hypothesis of this experiment would be the same that yeast extracts up-regulate gene expression during heat shock the same way bacteria cells up-regulate gene expression during heat shock. Adding a lower amount of protein extract will produce fewer bands, but will eliminate the cluster of bands seen in lanes 1 and 3 of the current experiment (Fig. 1). In lane 3 of figure 1, instead of having a cluster of bands around 64KD, there might only be 1 band present at 64KD. The experiment addresses a technical question because a small part of the procedure is getting changed in order to improve the results. The technical question the experiment proposes is: are their bands present when 5 ug of protein extract are added to each well? If the experiment supports the hypothesis, then the lane with HSP 30 gene at 42°C will have one band at approximately 64 KD rather than a cluster of bands. This means a clear and accurate conclusion can be drawn from the results. The presence of one band at around 64 KD will still conclude that the expression of the HSP 30 gene is greater at 42°C than 30°C. However, if a band is not present around 64KD, then the results would not support the hypothesis. Overall, the amount of protein extract added to each sample, before conducting gel electrophoresis, will pose a technical question that can improve the results and accurately conclude whether HSP 30 gene expression increases in heat