Team:CIDEB-UANL Mexico/labwork methods

From 2014hs.igem.org

(Difference between revisions)
Line 351: Line 351:
<p>Go to results</p>
<p>Go to results</p>
-
<p><b>Viability in salt (experiment 1)</b></p>
+
<br>
 +
<p><b><h2>Viability in salt (experiment 1)</h2></b></p>
 +
 
 +
<p>In the research on NhaS it was said that the gene gives certain resistance to salt, but the exact percentage of resistance is unknown. There were designed 3 experiments to know the viability in salt of the NhaS transformed bacteria, also in order to know the maximum concentration in which bacteria with the capture module can be exposed to later experiment with it and find out how much salt can capture.</p><p>
 +
As in the ligation on NhaS and pSB1C3 was obtain two types of bacteria (red and white) it us uncertain if the ligation occurred properly and why some are red and some are white, through the next three experiments it can be determined which type has the gene or if both of them have it. .</p><p>
 +
 
<p>Bacteria transformed with the capture plasmid were inoculated in Petri dishes with different concentrations of salt</p>
<p>Bacteria transformed with the capture plasmid were inoculated in Petri dishes with different concentrations of salt</p>

Revision as of 03:04, 15 June 2014

Methods Notebook Results Interpretations Conclusions Future Plans
iGEM CIDEB 2014 - Project

Methods

Experiments

In this section, it is described all the experiments designed and performed in order to prove experimentally our modules.

Capture Module

UV Experimentation

This experiment was designed in order to know if the UV promoter is working properly.

Procedure:

1. Inoculate by streak 2 Petri dishes with NhaS DNA in pSB1C3 Red bacteria

2. Repeat step 1 but with NhaS DNA in pSB1C3 White bacteria

3. Let them grow during one day in the incubator at 37°C

4. Expose the four Petri dishes to UV irradiation (302nm) during 2 hours

5. Take photos each 10 minutes and wait for results. (You can also take video during the 2 hours instead of the photos)

Go to results


Viability in salt (experiment 1)

In the research on NhaS it was said that the gene gives certain resistance to salt, but the exact percentage of resistance is unknown. There were designed 3 experiments to know the viability in salt of the NhaS transformed bacteria, also in order to know the maximum concentration in which bacteria with the capture module can be exposed to later experiment with it and find out how much salt can capture.

As in the ligation on NhaS and pSB1C3 was obtain two types of bacteria (red and white) it us uncertain if the ligation occurred properly and why some are red and some are white, through the next three experiments it can be determined which type has the gene or if both of them have it. .

Bacteria transformed with the capture plasmid were inoculated in Petri dishes with different concentrations of salt

IMG_0317

Image 1. All the 18 Petri dishes inoculated with NhaS Red in pSB1C3 of all the 9 used concentrations.


IMG_0317

Image 2. All the 18 Petri dishes inoculated with NhaS White in pSB1C3 of all the 9 used concentrations.


IMG_0317

Image 3. All the 18 Petri dishes inoculated with the Control bacteria of all the 9 used concentrations.

All the bacteria containing the NhaS in pSB1C3 (Red and White) survived to a 10% concentration of salt.

None of the control group lived in any concentration of salt.


IMG_0317

Image 4. Nine Petri dishes with the maximum concentration of salt (10%) used in this experiment. From up to bottom: NhaS Red in pSB1C3, NhaS White in pSB1C3 and the Control bacteria.


iGEM CIDEB 2014 - Footer

Retrieved from "http://2014hs.igem.org/Team:CIDEB-UANL_Mexico/labwork_methods"