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Team:Caltech/Week 3
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{{Caltech_iGEM_10| | {{Caltech_iGEM_10| | ||
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| - | + | {{:Team:Caltech/WeekTemplate}} | |
==Monday 6/28== | ==Monday 6/28== | ||
* Tested the success of the construct we made using digest and gel electrophoresis. | * Tested the success of the construct we made using digest and gel electrophoresis. | ||
| - | ** Digested ligation product from 6/26 with EcoRI and PstI. | + | ** Digested ligation product from [https://2010.igem.org/Team:Caltech/Week_2#Saturday_6.2F26 6/26] with EcoRI and PstI. |
** Ran digests on gel along with 1kb and 100kb molecular ladders. | ** Ran digests on gel along with 1kb and 100kb molecular ladders. | ||
* Gel result: No bands were visible other than those of the ladders. Amount of DNA used was most likely too low and thus the ligation product could not be confirmed. | * Gel result: No bands were visible other than those of the ladders. Amount of DNA used was most likely too low and thus the ligation product could not be confirmed. | ||
| + | [[Image:CIT6-28gel.jpg|thumb|Alt=An image of the test gel of our ligation product. The middle two lanes should have bands, but apparently have none. Lane 1: 100bp ladder, Lane 2: 3uL raw ligation product, Lane 3: 15uL ligation digest, Lane 4: 1kb ladder.|Image of a test gel of our third ligation product. Note that the middle two lanes are sadly empty.]] | ||
* Transformed additional BioBricks from 2010 Spring DNA Distribution into DH5alpha cells | * Transformed additional BioBricks from 2010 Spring DNA Distribution into DH5alpha cells | ||
** [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274100 K274100], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274200 K274200], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274002 K274002], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274003 K274003], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274004 K274004] | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274100 K274100], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274200 K274200], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274002 K274002], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274003 K274003], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274004 K274004] | ||
==Tuesday 6/29== | ==Tuesday 6/29== | ||
| + | * Transformation results: Transformation of the ampicillin-resistant BioBricks seemed to be successful. However, no bacterial growth was present in the plates and liquid cultures with tetracyclin present. This seems to indicate a problem with the tetracyclin used. | ||
| + | ** Of the plates with bacterial growth ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K274100 K274100 (red)], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274200 K274200 (orange)]), there was evidence of pigment production, indicating a successful transformation. | ||
| + | * Transformed 7 more bricks (pigments & our AND gate components): | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274110 K274110], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274210 K274210], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274003 K274003], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274004 K274004], [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0076 C0076], [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0077 C0077], [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0077 R0077], [http://partsregistry.org/wiki/index.php?title=Part:BBa_I1765001 I1765001] | ||
==Wednesday 6/30== | ==Wednesday 6/30== | ||
| + | * Transformation results: Only two plates did not have colonies on them. | ||
| + | ** The liquid cultures of the red and orange dyes, [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274110 K274110] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274210 K274210], seemed to be different colors than the other cultures. When centrifuged down, the cells showed signs of pigment production. | ||
| + | ** Both the colonies and the liquid culture of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274004 K274004 (light green)] were darkly colored. There was pigment production; however, the pigment did not appear to be a light green color. | ||
| + | * Prepared glycerol stocks. | ||
| + | * Retransformed three bricks and one new brick (GFP): | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_I765001 I765001] [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274004 K274004] [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0077 C0077] [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13504 I13504] | ||
==Thursday 7/1== | ==Thursday 7/1== | ||
| + | * Transformation results: All transformations were successful! There was no problem with the antibiotic used. | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274003 K274003 (dark green)] was a dark gray color - evidence of production of pigment. | ||
| + | ** The colonies of [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13504 I13504 (GFP reporter gene)] did not fluoresce under UV light. | ||
| + | * Performed minipreps and created glycerol stocks from the liquid cultures. | ||
| + | * Received requested bricks from the Registry of Standard Parts. We plated and created liquid cultures of the cells containing the bricks. | ||
| + | * Began digests of bricks according to the NEB BioBrick assembly kit protocol. | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_KI765001 KI765001] (upstream) | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_KI13504 KI13504] (downstream) | ||
| + | ** [http://partsregistry.org/Part:pSB1C3 pSB1C3] (plasmid backbone) | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 B0034] (upstream) | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274100 K274100] (downstream) | ||
| + | ** pBAD18 plasmid backbone (including an AraC operon) | ||
| + | * Began ligation reactions: | ||
| + | # [http://partsregistry.org/wiki/index.php?title=Part:BBa_KI765001 KI765001]/[http://partsregistry.org/wiki/index.php?title=Part:BBa_KI13504 KI13504]/[http://partsregistry.org/Part:pSB1C3 pSB1C3] | ||
| + | ::* Attaching the UV promoter to a GFP reporter will allow us to test the promoter. | ||
| + | # [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 B0034]/[http://partsregistry.org/wiki/index.php?title=Part:BBa_K274100 K274100]/pBAD18 | ||
| + | ::* To be used for a pigment-production experiment in minimal media. | ||
| + | * Transformed: | ||
| + | ** Ligation 1 & 2 products (above) | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0011 R0011] | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23119 J23119] | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0080 R0080] | ||
==Friday 7/2== | ==Friday 7/2== | ||
| + | Note: Friday is an Institute Holiday (which explains why we didn't do much today). | ||
| + | |||
| + | * Mini-preped & made freezer stocks of the following bricks: | ||
| + | ** [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0011 R0011], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23119 J23119], [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0080 R0080], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450 J04450], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K112806 K112806], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J13002 J13002], [http://partsregistry.org/wiki/index.php?title=Part:BBa_I737020 I737020], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K274001 K274001], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K215000 K215000], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J63010 J63010]. | ||
| + | ** Only Ligation 2 was successfully transformed. | ||
==Weekend 7/3-4== | ==Weekend 7/3-4== | ||
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}} | }} | ||
Latest revision as of 22:57, 16 July 2010
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People
|
 Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 | Week 15
Monday 6/28
Tuesday 6/29
Wednesday 6/30
Thursday 7/1
Friday 7/2Note: Friday is an Institute Holiday (which explains why we didn't do much today).
Weekend 7/3-4
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