Line 728: | Line 728: | ||
<br> | <br> | ||
<div style="text-indent:40px;"> | <div style="text-indent:40px;"> | ||
− | 1. Large qualitative screening of IAA-producing colonies at the same time to see if our constructs | + | 1. Large qualitative screening of IAA-producing colonies at the same time to see if our constructs<div style="text-indent:60px;"> are still functional in our E.coli/yeasts </div> </div> |
<div style="text-indent:40px;">2. Helps us o pic the right colonies for colony-PCR and GC-MS measurements </div> | <div style="text-indent:40px;">2. Helps us o pic the right colonies for colony-PCR and GC-MS measurements </div> | ||
<br> | <br> | ||
Line 737: | Line 737: | ||
1. Perchloric acid is highly corrosive and dangerous!!<div style="text-indent:80px;"> Always uses protective gear and work under a fume hood! </div> </div> | 1. Perchloric acid is highly corrosive and dangerous!!<div style="text-indent:80px;"> Always uses protective gear and work under a fume hood! </div> </div> | ||
<div style="text-indent:60px;"> | <div style="text-indent:60px;"> | ||
− | 2. Reagent will always be mixed together on the spot, FeCl3 stock solution is finished, acid will be taken from the chemicals sheld from the AG plant physiology (has been negotiated)</div> | + | 2. Reagent will always be mixed together on the spot, FeCl3 stock solution is finished, acid will <div style="text-indent:80px;"> be taken from the chemicals sheld from the AG plant physiology (has been negotiated)</div></div> |
<br> | <br> | ||
Line 748: | Line 748: | ||
<b>4. Assay conditions </b> | <b>4. Assay conditions </b> | ||
<div style="text-indent:40px;"> | <div style="text-indent:40px;"> | ||
− | 1. Plates were inoculated in a grid pattern and overlaid with an 82 mm-diameter disk of Nitrocellulose membranes</div> | + | 1. Plates were inoculated in a grid pattern and overlaid with an 82 mm-diameter disk of<div style="text-indent:60px;"> Nitrocellulose membranes</div> </div> |
<div style="text-indent:40px;"> | <div style="text-indent:40px;"> | ||
− | 2. Plates are overlaid with Nitrocellulose immediately after inoculation with toothpicks | + | 2. Plates are overlaid with Nitrocellulose immediately after inoculation with toothpicks<div style="text-indent:60px;"> |
− | After normal incubation (i.e. overnight) time, the membrane was removed and soaked in reagent (or reagent-saturated [2.5 mL] filter paper, here “Whatman grade 2” had best results), in glass chamber ( danger symbol and written information)</div> | + | After normal incubation (i.e. overnight) time, the membrane was removed and soaked in reagent <div style="text-indent:60px;">(or reagent-saturated [2.5 mL] filter paper, here “Whatman grade 2” had best results),<div style="text-indent:60px;"> in glass chamber ( danger symbol and written information)</div></div></div></div> |
<div style="text-indent:40px;"> | <div style="text-indent:40px;"> | ||
3. After 30 - 60 minutes, coloring reaction is finished and fading began </div> | 3. After 30 - 60 minutes, coloring reaction is finished and fading began </div> | ||
Line 757: | Line 757: | ||
4. Best results with colony sizes between 0.5 to 2 mm </div> | 4. Best results with colony sizes between 0.5 to 2 mm </div> | ||
<div style="text-indent:40px;"> | <div style="text-indent:40px;"> | ||
− | 5. Addition of Tryptophan greatly enhances color reaction but does not interfere with distinguishing IAA positive and negative colonies (yellow background and strong red to pink positives)</div> | + | 5. Addition of Tryptophan greatly enhances color reaction but does not interfere with<div style="text-indent:60px;"> distinguishing IAA positive and negative colonies (yellow background and strong red to pink<div style="text-indent:60px;"> positives)</div></div></div> |
<div style="text-indent:40px;"> | <div style="text-indent:40px;"> | ||
− | Other indolic compounds (i.e. indolepyruvic acid) are distinguishable by a more yellow-brownish color </div> | + | 6. Other indolic compounds (i.e. indolepyruvic acid) are distinguishable by a more yellow-brownish<div style="text-indent:60px;"> color </div></div> |
<br> | <br> | ||
<b> 5. Afterwards </b> | <b> 5. Afterwards </b> |
Revision as of 16:04, 29 October 2017
Our research work
We are describing our research work. Below you can find the protocols we used.
Protocols
Main sponsors: