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Virginia Commonwealth/14 August 2009
From 2009.igem.org
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*Because the concentrations were so low for J23113, J23116, and R0011, more frozen stocks will be grown up and Miniprepped. R0040, J23110 have more minipreps in stock in the freezer | *Because the concentrations were so low for J23113, J23116, and R0011, more frozen stocks will be grown up and Miniprepped. R0040, J23110 have more minipreps in stock in the freezer | ||
* Electrophoresis Gel Results | * Electrophoresis Gel Results | ||
| + | ** It is really difficult to see promoters from digestions when run on the gel | ||
[[Image:2009-08-14 digest 106,107,110,113,116,0011,0040,110mini.jpg|350px|thumb|none|From right to left: HyperLadder I, Low Molecular Weight Ladder, Digests: J23106, J23107, J23110, J23113, J23116, R0011, R0040, Miniprepped J23110]] | [[Image:2009-08-14 digest 106,107,110,113,116,0011,0040,110mini.jpg|350px|thumb|none|From right to left: HyperLadder I, Low Molecular Weight Ladder, Digests: J23106, J23107, J23110, J23113, J23116, R0011, R0040, Miniprepped J23110]] | ||
| Line 75: | Line 76: | ||
* Today I am running a gel on the purified PCR products to confirm the reaction. The gel is 1% agarose, running at 100 volts for 45 minutes. | * Today I am running a gel on the purified PCR products to confirm the reaction. The gel is 1% agarose, running at 100 volts for 45 minutes. | ||
| - | * | + | ''Maria and Afton'' |
| - | + | * Make Cryogenic stock of the overnight cultures J23100, J23101 | |
| + | * Miniprep J23100 and J23101 | ||
| + | * Take Spectrophotometry of J23110, J23113, J23116, R0011, R0040 | ||
| + | * Digest J23110, J23113, J23116, R0011, R0040 | ||
| + | * Run a gel on Digests: J23106, J23107, J23110, J23113, J23116, R0011, R0040, Miniprepped 110 | ||
| + | ** Miniprepped J23110 was chosen because it had the greatest amount of DNA at the time | ||
| + | * Ligate J23110, J23113, J23116, R0011, R0040 with pSB1C3 w/p1010, J06702 (RET) | ||
| + | ** pSB1C3 w/p1010, J06702 (RET) were digested and were run on a gel August 12 and was reused for the ligations done today | ||
| + | ** Ligations will be frozen until cuvettes come in | ||
| + | [[User:Trentay|Trentay]] 23:05, 17 August 2009 (UTC) | ||
---- | ---- | ||
====Wetlab==== | ====Wetlab==== | ||
| - | * | + | ''Maria and Afton'' |
| - | * | + | * Miniprep was done |
| + | * LB was made | ||
| + | * Spectrophotometry was done | ||
| + | * Digestion was done | ||
| + | ** Next time BSA and other BioBrick Assmbly Materials are aliquoted, they should be placed in smaller tubes, possibly PCR tubes, because it is very difficult to see the samples | ||
| + | * Electrophoresis Test was done | ||
| + | ** Smaller wells were used so more samples could be placed on one gel (15 wells instead of 8) | ||
| + | [[User:Trentay|Trentay]] 23:08, 17 August 2009 (UTC) | ||
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Contents |
Friday 14 August 2009
Results
- Gel did not show any results.
Maria and Afton
- Overnight cultures of J23100 and J23101
- J23100 was vaguely pink
- J23101 was slightly pink
- They should probably be left for another day to grow to see more expression
Spectrophotometry Measurements for Digestion
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
|---|---|---|---|---|---|---|
| J23110 | 0.037 | 0.027 | 1.378 | 37 | 74 | 2.74 |
| J23113 | 0.020 | 0.012 | 1.667 | 22 | 40 | 0.88 |
| J23116 | 0.015 | 0.010 | 1.500 | 27 | 30 | 0.81 |
| R0011 | 0.010 | 0.007 | 1.420 | 28 | 20 | 0.56 |
| R0040 | 0.009 | 0.007 | 1.286 | 24 | 18 | 0.432 |
- Because the concentrations were so low for J23113, J23116, and R0011, more frozen stocks will be grown up and Miniprepped. R0040, J23110 have more minipreps in stock in the freezer
- Electrophoresis Gel Results
- It is really difficult to see promoters from digestions when run on the gel
Trentay 22:05, 17 August 2009 (UTC)
Tasks
- Today I am running a gel on the purified PCR products to confirm the reaction. The gel is 1% agarose, running at 100 volts for 45 minutes.
Maria and Afton
- Make Cryogenic stock of the overnight cultures J23100, J23101
- Miniprep J23100 and J23101
- Take Spectrophotometry of J23110, J23113, J23116, R0011, R0040
- Digest J23110, J23113, J23116, R0011, R0040
- Run a gel on Digests: J23106, J23107, J23110, J23113, J23116, R0011, R0040, Miniprepped 110
- Miniprepped J23110 was chosen because it had the greatest amount of DNA at the time
- Ligate J23110, J23113, J23116, R0011, R0040 with pSB1C3 w/p1010, J06702 (RET)
- pSB1C3 w/p1010, J06702 (RET) were digested and were run on a gel August 12 and was reused for the ligations done today
- Ligations will be frozen until cuvettes come in
Trentay 23:05, 17 August 2009 (UTC)
Wetlab
Maria and Afton
- Miniprep was done
- LB was made
- Spectrophotometry was done
- Digestion was done
- Next time BSA and other BioBrick Assmbly Materials are aliquoted, they should be placed in smaller tubes, possibly PCR tubes, because it is very difficult to see the samples
- Electrophoresis Test was done
- Smaller wells were used so more samples could be placed on one gel (15 wells instead of 8)
Trentay 23:08, 17 August 2009 (UTC)
