Team:Caltech/Notebook/Degradation
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<font size="+2"><a name="6_22_12">June 22, 2012</a></font> | <font size="+2"><a name="6_22_12">June 22, 2012</a></font> | ||
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<br> Created a spreadsheet detailing various possible bacterial strains and plasmids to be used for complex polymer degradation | <br> Created a spreadsheet detailing various possible bacterial strains and plasmids to be used for complex polymer degradation | ||
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+ | <font size="+2"><a name="6_25_29_12">June 25 - 29, 2012</a></font> | ||
<br> Sent out requests for bacterial strains and plasmids based on spreadsheet | <br> Sent out requests for bacterial strains and plasmids based on spreadsheet | ||
<br> Emailed Dr. Jared Leadbetter asking if he has strains + protocol for lignocellulose plates | <br> Emailed Dr. Jared Leadbetter asking if he has strains + protocol for lignocellulose plates | ||
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<br> Emailed for samples of nitrocellulose, latex, and teflon membranes. | <br> Emailed for samples of nitrocellulose, latex, and teflon membranes. | ||
<br> Researched Z. mobilis. | <br> Researched Z. mobilis. | ||
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<a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | <a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | ||
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<font size="+2"><a name="7_2_6_12">July 2-6, 2012</a></font> | <font size="+2"><a name="7_2_6_12">July 2-6, 2012</a></font> | ||
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<br> Researched zymomonas mobilis, narrowed down focus to strain ZM4 | <br> Researched zymomonas mobilis, narrowed down focus to strain ZM4 | ||
<br> Gathered more information on protocols specific to that strain. | <br> Gathered more information on protocols specific to that strain. | ||
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<br> Made 20% glucose solution stock and RM plates for Z. mobilis | <br> Made 20% glucose solution stock and RM plates for Z. mobilis | ||
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<a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | <a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | ||
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<font size="+2"><a name="7_9_13_12">July 9-13, 2012</a></font> | <font size="+2"><a name="7_9_13_12">July 9-13, 2012</a></font> | ||
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<br> Ordered strains from the ATCC. | <br> Ordered strains from the ATCC. | ||
<br> Discussed Biolog plates idea | <br> Discussed Biolog plates idea | ||
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<br> Sent in request for oligos sequences for alpha-gluctosidase gene (can degrade maltose) | <br> Sent in request for oligos sequences for alpha-gluctosidase gene (can degrade maltose) | ||
<br> Made tetracycline plates | <br> Made tetracycline plates | ||
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<a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | <a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | ||
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<br> There was no growth on the pSB1T3 plates. I kept a liquid culture of the electroporated cells that were left over after plating the on the tetra plates, and the tube looked very cloudy. The liquid culture did not have any antibiotics, but the plates had tetracycline. So it is possible that the gene is broken or its the wrong part or something. We decided to get the tetra resistance gene from a different source on the registry plates. So, I transformed BBa_K274002 and BBa_K274003 into E. coli from 2012 Kit Plate 3 12B, 2012 Kit Plate 4 20D, and 2012 Kit Plate 3 22D. I was originally trying to use BBa_J04450 from Plate 1, well 7A. | <br> There was no growth on the pSB1T3 plates. I kept a liquid culture of the electroporated cells that were left over after plating the on the tetra plates, and the tube looked very cloudy. The liquid culture did not have any antibiotics, but the plates had tetracycline. So it is possible that the gene is broken or its the wrong part or something. We decided to get the tetra resistance gene from a different source on the registry plates. So, I transformed BBa_K274002 and BBa_K274003 into E. coli from 2012 Kit Plate 3 12B, 2012 Kit Plate 4 20D, and 2012 Kit Plate 3 22D. I was originally trying to use BBa_J04450 from Plate 1, well 7A. | ||
<br> I was able to liquid cultures of pMQ30, pMQ95, pMQ97, DKN79 (DH5alpha + pLAFR5), and DKN1005 (BW20427 [WM3064] + pLAFR5). There has been no growth on the Z. mobilis plates yet. There seems to be growth in the Z. mobilis liquid culture, but not enough to make a glycerol stock. I designed primers for switching out resistance on pMQ95 or pMQ97 and for getting the tet resistance gene from the plated electroporated cells. | <br> I was able to liquid cultures of pMQ30, pMQ95, pMQ97, DKN79 (DH5alpha + pLAFR5), and DKN1005 (BW20427 [WM3064] + pLAFR5). There has been no growth on the Z. mobilis plates yet. There seems to be growth in the Z. mobilis liquid culture, but not enough to make a glycerol stock. I designed primers for switching out resistance on pMQ95 or pMQ97 and for getting the tet resistance gene from the plated electroporated cells. | ||
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+ | <br> Growth has appeared on the Z. mobilis plates!! | ||
+ | <br> Cleaned out the lab space | ||
+ | <br> Made glycerol stock of Z. mobilis (one without gentamicin, one with gent) | ||
+ | <br> Made liquid cultures of DKN1005 (BW20427 [WM3064] + pLAFR5), part from 2012 registry plate 1 7A, and 2012 registry part from plate 4 20A | ||
+ | <br> Made glycerol stocks from liquid cultures of DKN1005 (BW20427 [WM3064] + pLAFR5) and part from plate 1 7A | ||
+ | <br> Made more LB + tetracycline plates | ||
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+ | <br> Z. mobilis liquid cultures currently growing: | ||
+ | <br> 1. original (only Rich Media) | ||
+ | <br> 2. RM + amp (taken from original) | ||
+ | <br> 3. RM + amp (taken from original) | ||
+ | <br> 4. RM + gent (taken from plate) | ||
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<a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | <a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | ||
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+ | <font size="+2"><a name="7_23_27_12">July 23-27, 2012</a></font> | ||
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+ | <a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> |
Revision as of 02:19, 21 July 2012
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Degradation Notebook
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