Team:IIT Madras/Results

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<html><b><font color="#000">Fig 8.1: </font></b><i>DH5a grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics.</i></html>
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<html><b><font color="#000">Fig 8.1: </font></b><i>a) DH5a grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics. This clearly shows that DH5a cannot grow in medium with antibiotics. b) RFP (pSB1C3) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics. c) CFP (pSB1A2) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics. d) RFP (pSB1C3) - CFP (pSB1A2) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics.</i></html>
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<html><b><font color="#000">Fig 8.2: </font></b><i>RFP (pSB1C3) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics.</i></html>
 
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<html><b><font color="#000">Fig 8.3: </font></b><i>CFP (pSB1A2) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics.</i></html>
 
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<html><b><font color="#000">Fig 8.4: </font></b><i>RFP (pSB1C3) - CFP (pSB1A2) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics.</i></html>
 
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<html><b><font color="#000">Fig 8.5: </font></b><i>All 4 strains in media without antibiotics.</i></html>
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<html><b><font color="#000">Fig 8.2: </font></b><i>All 4 strains in media without antibiotics. This graph clearly elucidates the fact that the cells which bear a plasmid tend to grow slower than those without it. The DH5a cells show considerably faster growth rate in the exponential phase compared to all the transformed cells. We wish to use this result along with the fact that cells lose plasmids in the absence of an selection pressure to retain it, to direct plasmid loss in a very specific manner, which is the key requirement to achieve a locking system.</i></html>
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<html><b><font color="#000">Fig 8.6: </font></b><i>All 4 strains in their corresponding antibiotic media - DH5a in Lb without antibiotics, RFP (pSB1C3) in LB with Chloramphenicol, CFP (pSB1A2) in LB with Ampicillin, RFP (pSB1C3) - CFP (pSB1A2) cells in LB with Chloramphenicol and Ampicillin.</i></html>
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<html><b><font color="#000">Fig 8.3: </font></b><i>All 4 strains in their corresponding antibiotic media - DH5a in Lb without antibiotics, RFP (pSB1C3) in LB with Chloramphenicol, CFP (pSB1A2) in LB with Ampicillin, RFP (pSB1C3) - CFP (pSB1A2) cells in LB with Chloramphenicol and Ampicillin.</i></html>
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<html><b><font color="#000">Fig 8.7: </font></b><i>log (OD600) vs Time plot for all 4 strains in media without antibiotics.</i></html>
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<html><b><font color="#000">Fig 8.4: </font></b><i>log (OD600) vs Time plot for all 4 strains in media without antibiotics.</i></html>
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<html><b><font color="#000">Fig 8.8: </font></b><i>log (OD600) vs Time plot for all 4 strains in their corresponding antibiotic media - DH5a in Lb without antibiotics, RFP (pSB1C3) in LB with Chloramphenicol, CFP (pSB1A2) in LB with Ampicillin, RFP (pSB1C3) - CFP (pSB1A2) cells in LB with Chloramphenicol and Ampicillin.</i></html>
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<html><b><font color="#000">Fig 8.5: </font></b><i>log (OD600) vs Time plot for all 4 strains in their corresponding antibiotic media - DH5a in Lb without antibiotics, RFP (pSB1C3) in LB with Chloramphenicol, CFP (pSB1A2) in LB with Ampicillin, RFP (pSB1C3) - CFP (pSB1A2) cells in LB with Chloramphenicol and Ampicillin.</i></html>
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For updated results, keep checking <a href="http://openwetware.org/wiki/IGEM:Indian_Institue_of_Technology_Madras/2009/Notebook/PLASMID_-_Plasmid_Locking_Assembly_for_Sustaining_Multiple_Insert_DNA/2009/10/31">here.</a>
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For updated results, keep checking <a href="https://2009.igem.org/User:Abdul">here.</a>
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===Fluorescent Imaging===
 
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===Fluorescent Imaging===
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<html><p align="center"><html><b><font color="#000">Fig 9: </font></b><i>Co-transformed cells with both CFP(1A2) and RFP(1C3)seen under bright field, red filter and cyan filter.</i></html>
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For updated results, keep checking <a href="http://openwetware.org/wiki/IGEM:Indian_Institue_of_Technology_Madras/2009/Notebook/PLASMID_-_Plasmid_Locking_Assembly_for_Sustaining_Multiple_Insert_DNA/2009/10/31">here.</a>
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For updated results, keep checking <a href="https://2009.igem.org/User:Abdul">here.</a>
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{{:Team:IITM/footer}}

Latest revision as of 01:50, 22 October 2009

Indian Institute of Technology,Madras

IIT Madras













Contents

Results

Comparing the differences in the growth rates of cells with and without plasmids in various media


Growth curves


Clubbed.jpg
Fig 8.1: a) DH5a grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics. This clearly shows that DH5a cannot grow in medium with antibiotics. b) RFP (pSB1C3) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics. c) CFP (pSB1A2) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics. d) RFP (pSB1C3) - CFP (pSB1A2) contanining cells grown in LB containing no antibiotic, LB containing Ampicillin, LB containing Chloramphenicol, LB containing both antibiotics.



Noaball4.jpg
Fig 8.2: All 4 strains in media without antibiotics. This graph clearly elucidates the fact that the cells which bear a plasmid tend to grow slower than those without it. The DH5a cells show considerably faster growth rate in the exponential phase compared to all the transformed cells. We wish to use this result along with the fact that cells lose plasmids in the absence of an selection pressure to retain it, to direct plasmid loss in a very specific manner, which is the key requirement to achieve a locking system.



Corraball4.jpg
Fig 8.3: All 4 strains in their corresponding antibiotic media - DH5a in Lb without antibiotics, RFP (pSB1C3) in LB with Chloramphenicol, CFP (pSB1A2) in LB with Ampicillin, RFP (pSB1C3) - CFP (pSB1A2) cells in LB with Chloramphenicol and Ampicillin.



Logall4noab.jpg
Fig 8.4: log (OD600) vs Time plot for all 4 strains in media without antibiotics.



Logcorrab.jpg
Fig 8.5: log (OD600) vs Time plot for all 4 strains in their corresponding antibiotic media - DH5a in Lb without antibiotics, RFP (pSB1C3) in LB with Chloramphenicol, CFP (pSB1A2) in LB with Ampicillin, RFP (pSB1C3) - CFP (pSB1A2) cells in LB with Chloramphenicol and Ampicillin.

Modeling

For updated results, keep checking here.

Fluorescent Imaging


Fluorescence merge pic.jpg

Fig 9: Co-transformed cells with both CFP(1A2) and RFP(1C3)seen under bright field, red filter and cyan filter.





For updated results, keep checking here.


 

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