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Team:Caltech/Week 2
From 2011.igem.org
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<p>Lab walkthrough<br/> | <p>Lab walkthrough<br/> | ||
Setting up lab, stock supplies, preparation of LB-amp plates<br/> | Setting up lab, stock supplies, preparation of LB-amp plates<br/> | ||
| - | Test extraction of DNA from BioBrick wells into cloning plates<br/> | + | Test extraction of DNA from BioBrick wells into cloning plates ([http://partsregistry.org/Part:BBa_K123000 K123000], [http://partsregistry.org/Part:BBa_K123001 K123001], [http://partsregistry.org/Part:BBa_K123002 K123002], [http://partsregistry.org/Part:BBa_K123003 K123003])<br/> |
Initial enrichment cultures of collected samples on minimal media with BPA or 17a-ethynylestradiol</p> | Initial enrichment cultures of collected samples on minimal media with BPA or 17a-ethynylestradiol</p> | ||
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<table border="1"> | <table border="1"> | ||
<tr> | <tr> | ||
| - | <th> | + | <th>Sample</th> |
<th>Concentration (ng/µl)</th> | <th>Concentration (ng/µl)</th> | ||
</tr> | </tr> | ||
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Locations 3 and 8 were omitted from the MoBio Powermax Soil Extraction procedure because they were liquid samples. | Locations 3 and 8 were omitted from the MoBio Powermax Soil Extraction procedure because they were liquid samples. | ||
== June 22 == | == June 22 == | ||
| - | <p>Miniprep and sequencing of selected BioBricks ( | + | <p>Miniprep and sequencing of selected BioBricks (BBa_K123000, BBa_K123001, BBa_K123002, BBa_K123003)<br/> |
Preparation of competent cells</p> | Preparation of competent cells</p> | ||
| - | + | Results of OD measurements: | |
<table border="1"> | <table border="1"> | ||
<tr> | <tr> | ||
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==June 23== | ==June 23== | ||
| - | <p> | + | <p>Transformation of T7 polymerase ([http://partsregistry.org/Part:BBa_K145001 K145001]), mCherry ([http://partsregistry.org/Part:BBa_J06702 J06702]]), Lac Promoter ([http://partsregistry.org/Part:BBa_R0010 R0010]]), and double terminator ([http://partsregistry.org/Part:BBa_B0014 B0014] and [http://partsregistry.org/Part:BBa_B0015 B0015]) biobricks for test sequences for BisA and BisB degradation genes ([http://partsregistry.org/Part:BBa_K123000 K123000] and [http://partsregistry.org/Part:BBa_K123001 K123001])<br/> |
| - | + | Retested competent cells made June 22nd with [http://partsregistry.org/Part:BBa_K123001 K123001] and pUC<br/> | |
| - | + | Transfered 0.5 mL aliquots of BPA and 5mL 17a-ethynylestradiol cultures from June 20 to new tubes of minimal media<br/> | |
| - | + | Designed primers for test sequence of BisdA (pNT001)<br/> | |
| - | + | Designed primers to continue sequencing of human ER ([http://partsregistry.org/Part:BBa_K123003 K123003])<br/><br/> | |
| - | + | ||
We determined that sequences for [http://partsregistry.org/Part:BBa_K123000 K123000] and [http://partsregistry.org/Part:BBa_K123001 K123001] have correct protein coding but different codons than in the sequences shown online </p> | We determined that sequences for [http://partsregistry.org/Part:BBa_K123000 K123000] and [http://partsregistry.org/Part:BBa_K123001 K123001] have correct protein coding but different codons than in the sequences shown online </p> | ||
==June 24== | ==June 24== | ||
| - | <p> | + | <p>Transformation of Tet repressible promoter [http://partsregistry.org/Part:BBa_R0040 R0040]<br/> |
| - | + | Preparation of LB, LB (no antibiotic) plates, and SOC media<br/> | |
| - | + | Transfer of 5 mL DDT and nonylphenol cultures from June 21 to new tubes of minimal media <br/> | |
| - | Transfer 5 mL DDT and nonylphenol cultures from June 21 to new tubes of minimal media | + | Design of reverse primer for continued sequencing of human ER ([http://partsregistry.org/Part:BBa_K123003 K123003]) <br/> |
| - | + | ||
| - | Design reverse primer for continued sequencing of human ER ([http://partsregistry.org/Part:BBa_K123003 K123003]) <br/> | + | |
Update Wiki with protocols we have been using<br/><br/> | Update Wiki with protocols we have been using<br/><br/> | ||
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We determined that our competent cells are functional have a cfu/ug DNA of 6.6 x 10^6.</p> | We determined that our competent cells are functional have a cfu/ug DNA of 6.6 x 10^6.</p> | ||
Latest revision as of 21:56, 1 August 2011
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Project |
June 19Collection of soil and water samples from LA River June 20Lab walkthrough June 21MoBio Powermax Soil kit extraction of DNA from collected samples ResultsMoBio Extraction
Locations 3 and 8 were omitted from the MoBio Powermax Soil Extraction procedure because they were liquid samples. June 22Miniprep and sequencing of selected BioBricks (BBa_K123000, BBa_K123001, BBa_K123002, BBa_K123003) Results of OD measurements:
June 23Transformation of T7 polymerase ([http://partsregistry.org/Part:BBa_K145001 K145001]), mCherry ([http://partsregistry.org/Part:BBa_J06702 J06702]]), Lac Promoter ([http://partsregistry.org/Part:BBa_R0010 R0010]]), and double terminator ([http://partsregistry.org/Part:BBa_B0014 B0014] and [http://partsregistry.org/Part:BBa_B0015 B0015]) biobricks for test sequences for BisA and BisB degradation genes ([http://partsregistry.org/Part:BBa_K123000 K123000] and [http://partsregistry.org/Part:BBa_K123001 K123001]) June 24Transformation of Tet repressible promoter [http://partsregistry.org/Part:BBa_R0040 R0040] June 25Take out plates from June 24
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