Difference between revisions of "Team:Manchester/Demonstrate"
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<div class="col-md-12 text-justify"> | <div class="col-md-12 text-justify"> | ||
| − | <p><font color="#111"> | + | <p><font color="#111">A control DAPI stain was performed along side Medium promoter tag expression to inspect the DAPI distribution in the absence of polyphosphate and whether the staining procedure interfered with mCherry distribution.<br> |
| + | <br> | ||
| + | <center> | ||
| + | <img src="https://static.igem.org/mediawiki/2017/a/a9/MidDAPIcontrol.png" width="800" height="317"> | ||
| + | </center> | ||
| + | <br> | ||
| + | <div class="col-md-12 text-justify"> | ||
| + | <p><font color="#111">As shown, the distribution of both mCherry and DAPI were homogeneous with no obvious clumping or accumulation. This suggested that if our construct was working properly, we would see an accumulation of fluorescent signal for both mCherry and DAPI in the same place within the cell.<br> | ||
| + | <br> | ||
| + | So in line with this, we DAPI stained a 24h induction of Low+EutSMNLK+PPK:<br> | ||
| + | <br> | ||
| + | <center> | ||
| + | <img src="https://static.igem.org/mediawiki/2017/d/d0/SMNLKDAPI1.png" width="300" height="300"> | ||
| + | <img src="https://static.igem.org/mediawiki/2017/b/b5/SMNLKDAPI2.png" width="300" height="300"> | ||
| + | <img src="https://static.igem.org/mediawiki/2017/f/fe/SMNLKDAPI3.png" width="300" height="300"> | ||
| + | </center> | ||
| + | <br> | ||
| + | <div class="col-md-12 text-justify"> | ||
| + | <p><font color="#111">As shown, there is a heterogeneous distribution of fluorescence within the cells for both signals and they are approximately in the same areas. This heterogeneous distribution of DAPI indicates the presence of polyphosphate and proves the activity of our PPK along with its successful localization into our BMC. The localisation can be determined using the physical location of both fluorescence signals within the cell.<br> | ||
| + | <br> | ||
| + | </font></p> | ||
Revision as of 19:28, 29 October 2017
Demonstrate
Microscopy
A control DAPI stain was performed along side Medium promoter tag expression to inspect the DAPI distribution in the absence of polyphosphate and whether the staining procedure interfered with mCherry distribution.
As shown, the distribution of both mCherry and DAPI were homogeneous with no obvious clumping or accumulation. This suggested that if our construct was working properly, we would see an accumulation of fluorescent signal for both mCherry and DAPI in the same place within the cell.
So in line with this, we DAPI stained a 24h induction of Low+EutSMNLK+PPK:
As shown, there is a heterogeneous distribution of fluorescence within the cells for both signals and they are approximately in the same areas. This heterogeneous distribution of DAPI indicates the presence of polyphosphate and proves the activity of our PPK along with its successful localization into our BMC. The localisation can be determined using the physical location of both fluorescence signals within the cell.
