Difference between revisions of "Team:Vanderbilt/Experiments/LCMS"
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<b> Summary </b> | <b> Summary </b> | ||
<p> Liquid chromatography--mass spectrometry (LC-MS) is the current standard for measuring DNA oxidation due to its high sensitivity and specificity. We experimented with methods to analyze levels of oxidation in our gene samples, and achieved reliable detection of hydrolyzed bases. We could detect 8-oxoguanine signal in our analytical standard, and characterized four discrete protocols for in vitro oxidation at a range of doses (hydrogen peroxide, methylene blue/light, UV irradiation, Mg). Although signal was often low, we have begun to compare our optimized sequences with improved oxidation methods.</p> | <p> Liquid chromatography--mass spectrometry (LC-MS) is the current standard for measuring DNA oxidation due to its high sensitivity and specificity. We experimented with methods to analyze levels of oxidation in our gene samples, and achieved reliable detection of hydrolyzed bases. We could detect 8-oxoguanine signal in our analytical standard, and characterized four discrete protocols for in vitro oxidation at a range of doses (hydrogen peroxide, methylene blue/light, UV irradiation, Mg). Although signal was often low, we have begun to compare our optimized sequences with improved oxidation methods.</p> | ||
| − | <p> With regard to 8-oxoG standard detection, we were compelled to make a number of modifications to our protocol to detect our compound within a triple quadrupole mass analyzer. Xcalibur software was used to predict fragment masses that we eventually concluded could not be detected in our sample. Fortunately, a number of predictive modifications to our model demonstrated our compound could be detected in our sample and uncovered the corresponding elution time. </p> | + | <p> With regard to 8-oxoG standard detection, we were compelled to make a number of modifications to our protocol to detect our compound within a triple quadrupole mass analyzer. Xcalibur software was used to predict fragment masses that we eventually concluded could not be detected in our sample. Fortunately, a number of predictive modifications to our model demonstrated our compound could be detected in our sample and uncovered the corresponding elution time. <br> |
| + | <img src="https://static.igem.org/mediawiki/2016/4/4f/Vu16_figure10.png" alt="project" height=“1215" width=“2160”/> <br> | ||
| + | <img src="https://static.igem.org/mediawiki/2016/a/a6/Vu16_figure11.png" alt="project" height=“1215" width=“2160”/> | ||
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Revision as of 03:50, 20 October 2016
Mass-Spectroscopy
SummaryLiquid chromatography--mass spectrometry (LC-MS) is the current standard for measuring DNA oxidation due to its high sensitivity and specificity. We experimented with methods to analyze levels of oxidation in our gene samples, and achieved reliable detection of hydrolyzed bases. We could detect 8-oxoguanine signal in our analytical standard, and characterized four discrete protocols for in vitro oxidation at a range of doses (hydrogen peroxide, methylene blue/light, UV irradiation, Mg). Although signal was often low, we have begun to compare our optimized sequences with improved oxidation methods.
With regard to 8-oxoG standard detection, we were compelled to make a number of modifications to our protocol to detect our compound within a triple quadrupole mass analyzer. Xcalibur software was used to predict fragment masses that we eventually concluded could not be detected in our sample. Fortunately, a number of predictive modifications to our model demonstrated our compound could be detected in our sample and uncovered the corresponding elution time.
