Difference between revisions of "Part:BBa R0077:Experience"

(→‎User Reviews)
(→‎User Reviews)
 
Line 13: Line 13:
 
|width='10%'|
 
|width='10%'|
 
<partinfo>BBa_R0077 AddReview number</partinfo>
 
<partinfo>BBa_R0077 AddReview number</partinfo>
−
<I>Username</I>
 
 
|width='60%' valign='top'|
 
|width='60%' valign='top'|
 
Enter the review inofrmation here.
 
Enter the review inofrmation here.
Line 19: Line 18:
 
<!-- End of the user review template -->
 
<!-- End of the user review template -->
 
<!-- DON'T DELETE --><partinfo>BBa_R0077 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_R0077 EndReviews</partinfo>
 +
 +
 +
{|width='80%' style='border:1px solid gray'
 +
|-
 +
|width='10%'|
 +
<partinfo>BBa_R0079 AddReview number</partinfo>
 +
<I>Peking_S iGEM 2011</I>
 +
|width='60%' valign='top'|
 +
 
First, we induced generator cell with 10-3M IPTG for 16 hours, then centrifuged cells and collect the supernatant. We use this supernatant as original 3OH-C14:1-HSL solution，then we diluted it to 10-7,  5×10-7,  10-6, 5×10-6, 10-5, 6×10-5, 8×10-5, 10-4, 2×10-4, 4×10-4, 7×10-4, 10-3, 3×10-3, 6×10-3, 8×10-3, 10-2 of original solution concentration respectively. Using these diluted supernatants to induce receiver cells (OD600 0.4) at 37℃ for 3 hours, then centrifuged cells and resuspended in phosphate buffer solution (PBS). The GFP fluorescence of each culture was obtained using enzyme-labeled assay. The result is shown in figure 1.
 
First, we induced generator cell with 10-3M IPTG for 16 hours, then centrifuged cells and collect the supernatant. We use this supernatant as original 3OH-C14:1-HSL solution，then we diluted it to 10-7,  5×10-7,  10-6, 5×10-6, 10-5, 6×10-5, 8×10-5, 10-4, 2×10-4, 4×10-4, 7×10-4, 10-3, 3×10-3, 6×10-3, 8×10-3, 10-2 of original solution concentration respectively. Using these diluted supernatants to induce receiver cells (OD600 0.4) at 37℃ for 3 hours, then centrifuged cells and resuspended in phosphate buffer solution (PBS). The GFP fluorescence of each culture was obtained using enzyme-labeled assay. The result is shown in figure 1.
  
Line 35: Line 43:
 
<center>[[Image:LM12a.png|600px]]</center>
 
<center>[[Image:LM12a.png|600px]]</center>
 
''Figure 3. Cin receiver system cell distribution of GFP fluorescence intensity.''
 
''Figure 3. Cin receiver system cell distribution of GFP fluorescence intensity.''
 +
|};

Latest revision as of 19:59, 18 October 2011

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_R0077

User Reviews

UNIQ94fc63a32b3e521f-partinfo-00000000-QINU UNIQ94fc63a32b3e521f-partinfo-00000001-QINU


No review score entered. Peking_S iGEM 2011

First, we induced generator cell with 10-3M IPTG for 16 hours, then centrifuged cells and collect the supernatant. We use this supernatant as original 3OH-C14:1-HSL solution，then we diluted it to 10-7, 5×10-7, 10-6, 5×10-6, 10-5, 6×10-5, 8×10-5, 10-4, 2×10-4, 4×10-4, 7×10-4, 10-3, 3×10-3, 6×10-3, 8×10-3, 10-2 of original solution concentration respectively. Using these diluted supernatants to induce receiver cells (OD600 0.4) at 37℃ for 3 hours, then centrifuged cells and resuspended in phosphate buffer solution (PBS). The GFP fluorescence of each culture was obtained using enzyme-labeled assay. The result is shown in figure 1.

Refigure1.png

Figure 1. Dose Response Curve. Vertical axis RFU/OD represents GFP expression measured through enzyme-labeled assay and the Horizontal axis represent different relative dilution concentration of 3OH-C14:1-HSL. Data was fitted with Hill function.

Time Dependence

We induced E.coli with receiver plasmid using supernatant diluted 10-2 times, and measure OD and absorbance of GFP every 15 minutes.

LM9.png

Figure 2. Time response curve of cin receiver system. We measured RFU and OD600 every 15 minutes and experiment performed in 3 hours.

Coordinatiblity

LM12a.png

Figure 3. Cin receiver system cell distribution of GFP fluorescence intensity.

;