Virginia Commonwealth/14 August 2009
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* 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) | ||
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====Wetlab==== | ====Wetlab==== | ||
* 1 | * 1 | ||
* 2 | * 2 |
Revision as of 23:05, 17 August 2009
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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
- 1
- 2