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<td>Cream/Light Green</td> | <td>Cream/Light Green</td> | ||
<td>Circular or Irregular form (occasionally rhizoid), Raised elevation, Shiny</td> | <td>Circular or Irregular form (occasionally rhizoid), Raised elevation, Shiny</td> | ||
− | <td>Colonies took on a different morphology depending on how the media was | + | <td>Colonies took on a different morphology depending on how the media was inoculated; stab-inoculation lead to rhizoid form while spreading leads to circular/irregular form</td> |
</tr> | </tr> | ||
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<img src="https://static.igem.org/mediawiki/2018/b/be/T--Newcastle--AllPreservePlatesNew.png"> | <img src="https://static.igem.org/mediawiki/2018/b/be/T--Newcastle--AllPreservePlatesNew.png"> | ||
<p> </p> | <p> </p> | ||
− | <font size="2">Figure 1: Observations of bacterial preservation plates. a) <i>A. caulinodans</i> colonies grown on 1 % Yeast Extract Broth agar after incubation at 30 °C for 56 hours. Plates were inoculated via streaking. b) <i>A. brasilense</i> colonies grown on 1 % LB after incubation at 37 °C for 16 hours. Plate inoculated via streaking. c) <i>H. seropedicae</i> colonies showing circular growth on 1 % LB agar after incubation at 30 °C for 24 hours. Plates were inoculated via streaking. d) <i>H. seropedicae</i> colonies showing rhizoid growth on 1 % LB agar after incubation at 30 °C for 24 hours. Plates were stab- | + | <font size="2">Figure 1: Observations of bacterial preservation plates. a) <i>A. caulinodans</i> colonies grown on 1 % Yeast Extract Broth agar after incubation at 30 °C for 56 hours. Plates were inoculated via streaking. b) <i>A. brasilense</i> colonies grown on 1 % LB after incubation at 37 °C for 16 hours. Plate inoculated via streaking. c) <i>H. seropedicae</i> colonies showing circular growth on 1 % LB agar after incubation at 30 °C for 24 hours. Plates were inoculated via streaking. d) <i>H. seropedicae</i> colonies showing rhizoid growth on 1 % LB agar after incubation at 30 °C for 24 hours. Plates were stab-inoculated. </font> |
</div> | </div> | ||
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<p style="font-size:medium">The response index, developed by Pham and Parkinson [4], accounts for a ratio between the edge of the colony nearest the chemoattractant source and the edge furthest from the same source. This ratio is then used to determine if there has been positive chemotaxis (RI > 0.52), no effect (RI = 0.48-0.52) or negative chemotaxis (RI < 0.48).</p> | <p style="font-size:medium">The response index, developed by Pham and Parkinson [4], accounts for a ratio between the edge of the colony nearest the chemoattractant source and the edge furthest from the same source. This ratio is then used to determine if there has been positive chemotaxis (RI > 0.52), no effect (RI = 0.48-0.52) or negative chemotaxis (RI < 0.48).</p> | ||
<p style="font-size:medium">Results (Table 6) indicated that both <i>A. brasilense</i> and <i>H. seropedicae</i> experienced positive chemotaxis towards 50 μM between distances of 5-25 mm and 5-10 mm respectively. As such, further investigation utilised the distance that corresponded with the greatest RI value (15 mm and 10 mm respectively). For <i>H. seropedicae</i>, the colonies nearer the centre line again showed more constricted halos which may indicate that the naringenin concentration may still be too high. The response index of the control for all species at 5 mm was < 0.48, suggesting chemorepulsion. This was anticipated as the control contains ethanol which possesses known antimicrobial properties and is commonly used to disinfect lab equipment.</p> | <p style="font-size:medium">Results (Table 6) indicated that both <i>A. brasilense</i> and <i>H. seropedicae</i> experienced positive chemotaxis towards 50 μM between distances of 5-25 mm and 5-10 mm respectively. As such, further investigation utilised the distance that corresponded with the greatest RI value (15 mm and 10 mm respectively). For <i>H. seropedicae</i>, the colonies nearer the centre line again showed more constricted halos which may indicate that the naringenin concentration may still be too high. The response index of the control for all species at 5 mm was < 0.48, suggesting chemorepulsion. This was anticipated as the control contains ethanol which possesses known antimicrobial properties and is commonly used to disinfect lab equipment.</p> | ||
− | <font size="2">Table 6: Average Response Index and standard error of <i>A. caulinodans</i>, <i>A. brasilense</i>, <i>H. seropedicae</i> and <i>E. coli</i> colonies grown on 0.25 % Minimal A Salt agar containing a gradient of either 100 µM naringenin or 1.5 % ethanol (control). RI = D1/(D1+D2) in which D1 represents distance between colony edge nearest chemical source to site of inoculation whilst D2 represents distance between colony edge furthest from chemical source to site of | + | <font size="2">Table 6: Average Response Index and standard error of <i>A. caulinodans</i>, <i>A. brasilense</i>, <i>H. seropedicae</i> and <i>E. coli</i> colonies grown on 0.25 % Minimal A Salt agar containing a gradient of either 100 µM naringenin or 1.5 % ethanol (control). RI = D1/(D1+D2) in which D1 represents distance between colony edge nearest chemical source to site of inoculation whilst D2 represents distance between colony edge furthest from chemical source to site of inoculation [4]. Bacteria were inoculated 15 mm (<i>A. brasilense</i> and <i>E. coli</i>) or 10 mm (<i>A. caulinodans</i> and <i>H. seropedicae</i>) from naringenin source and incubated at 30 ˚C.</font> |
<table id="protocols"> | <table id="protocols"> | ||
<thead> | <thead> |
Revision as of 00:29, 17 October 2018