Difference between revisions of "Team:UMaryland/Results"
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| − | <p style="font-size: | + | <p style="font-size:48px;text-align:center;font-family:Verdana, Geneva, sans-serif;"><b>Fluorescence Studies</b> |
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| + | <p style="font-size:32px;text-align:center;font-family:Verdana, Geneva, sans-serif;"><b>Analysis of Fluorescence Loss</b> | ||
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| + | <p><img src ="https://static.igem.org/mediawiki/2015/thumb/f/f2/UMaryland_Group_B_090615.JPG/800px-UMaryland_Group_B_090615.JPG" height="333px" width="500px"></p> | ||
<p style = "font-size:18px">Over time, colonies appear to stop producing RFP.</p> | <p style = "font-size:18px">Over time, colonies appear to stop producing RFP.</p> | ||
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| − | <p><img src = "https://static.igem.org/mediawiki/2015/2/26/UMaryland_Group_A_090615.JPG" | + | <p><img src = "https://static.igem.org/mediawiki/2015/2/26/UMaryland_Group_A_090615.JPG" height="333px" width="500px"></p> |
<p style = "font-size:18px"> This effect occurs even when the culture is under chloramphenicol pressure.</p> | <p style = "font-size:18px"> This effect occurs even when the culture is under chloramphenicol pressure.</p> | ||
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| − | <p><img src = "https://static.igem.org/mediawiki/2015/thumb/b/bd/UMaryland_Group_D_090615.JPG/800px-UMaryland_Group_D_090615.JPG"></p> | + | <div style="float:clear;"> |
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| + | <p><img src = "https://static.igem.org/mediawiki/2015/thumb/b/bd/UMaryland_Group_D_090615.JPG/800px-UMaryland_Group_D_090615.JPG"height="333px" width="500px"></p> | ||
<p style = "font-size:18px"> This effect occurs much more slowly when the Hok-Sok cassette is adjacent to the RFP construct.</p> | <p style = "font-size:18px"> This effect occurs much more slowly when the Hok-Sok cassette is adjacent to the RFP construct.</p> | ||
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| − | <p><img src= "https://static.igem.org/mediawiki/2015/thumb/9/92/UMaryland_Group_E_090615.JPG/800px-UMaryland_Group_E_090615.JPG"></p> | + | <p><img src= "https://static.igem.org/mediawiki/2015/thumb/9/92/UMaryland_Group_E_090615.JPG/800px-UMaryland_Group_E_090615.JPG"height="333px" width="500px"></p> |
<p style = "font-size:18px">With or without added chloramphenicol, Hok-Sok appears to be able to maintain a relatively constant level of fluorescence.</p> | <p style = "font-size:18px">With or without added chloramphenicol, Hok-Sok appears to be able to maintain a relatively constant level of fluorescence.</p> | ||
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| − | <p style="font-size: | + | <p style="font-size:48px;text-align:center;font-family:Verdana, Geneva, sans-serif;">Future Plans |
<p style="font-size:32px;text-align:center;font-family:Verdana, Geneva, sans-serif;">Considerations | <p style="font-size:32px;text-align:center;font-family:Verdana, Geneva, sans-serif;">Considerations | ||
<p style = "font-size:24px">PCR of Hok-Sok Construct</p> | <p style = "font-size:24px">PCR of Hok-Sok Construct</p> | ||
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<p style = "font-size:18px">Amplification of the Hok-Sok cassette is difficult due to the high inherent secondary structure in the construct. Hok ssDNA is capable of naturally folding into a stable secondary structure, resulting in early terminations and other side products. We recommend that a reaction buffer suitable for difficult templates be used, such as Phusion GC buffer + added DMSO.</p> | <p style = "font-size:18px">Amplification of the Hok-Sok cassette is difficult due to the high inherent secondary structure in the construct. Hok ssDNA is capable of naturally folding into a stable secondary structure, resulting in early terminations and other side products. We recommend that a reaction buffer suitable for difficult templates be used, such as Phusion GC buffer + added DMSO.</p> | ||
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<p style = "font-size:24px">Measuring OD of RFP Cultures</p> | <p style = "font-size:24px">Measuring OD of RFP Cultures</p> | ||
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<p style = "font-size:18px">Due to the close proximity of the emission wavelength of RFP (584 nm) and the classical absorbance wavelength for measuring cell density (600 nm), it is difficult to accurately determine the cell density of cultures that are expressing RFP. Given more time to calibrate our testing measurements, we would either have used an alternative wavelength for measuring OD (>600 nm), used a hemocytometer as an alternate counting method, or switched to GFP as an alternative fluorescent marker whose emission wavelength differs from 600 nm by a greater amount.</p> | <p style = "font-size:18px">Due to the close proximity of the emission wavelength of RFP (584 nm) and the classical absorbance wavelength for measuring cell density (600 nm), it is difficult to accurately determine the cell density of cultures that are expressing RFP. Given more time to calibrate our testing measurements, we would either have used an alternative wavelength for measuring OD (>600 nm), used a hemocytometer as an alternate counting method, or switched to GFP as an alternative fluorescent marker whose emission wavelength differs from 600 nm by a greater amount.</p> | ||
Revision as of 05:50, 18 September 2015
