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TUDelft/20 August 2008
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(New page: {{Template:TUDelftiGEM2008}} {{Template:TUDelftiGEM2008_menu_home}} <div id="tudelft_content"> =20th August= ==Midiprep== All the plasmids from live stabs have been midiprepped. The ge...) |
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| + | {{Template:TUDelftiGEM2008_calendar}} | ||
| - | + | =20th August 2008= | |
| - | = | + | ==Testing the Primers (part 2)== |
| + | Today another PCR was set up, again using the mastermix and composition also used for the first experiment (18th of August). Only this time the ''S. cerevisiae'' genes and cDNA were used. The cDNA contained 2ng/ul and the genes are: ERG8, ERG12, ERG13, HMG2 and MVD1. PCR program (with a 6ºC gradient this time) is as depicted below:<br> | ||
| + | 1. 5' @ 95ºC<br> | ||
| + | 2. 1' @ 95ºC<br> | ||
| + | 3. 1' @ 53.7ºC (gradient of 6C divided over 12 steps)<br> | ||
| + | 4. 1' @ 72ºC<br> | ||
| + | 5. repeat steps 2-4 29x (total of 30 cycles)<br> | ||
| + | 6. 5' @ 72ºC<br> | ||
| + | 7. forever @ 4ºC (PCR can be stopped and stored in the fridge at any time from this point on)<br> | ||
| - | + | This time, the products were put directly on gel. The expected sizes are: ERG8 - ~1400bp, ERG12 - ~1300bp, ERG13 - ~1500bp, MVD1 - ~1200bp and HMG2 - ~3000bp. As there was no visible result on the ERGs gel, only the MVD1/HMG2 gel is shown: | |
| - | + | ||
| + | [[Image:TUDelftgel3.jpg|thumb|center|MVD1/HMG2 gel]] | ||
| - | + | ==Midiprep== | |
| - | + | All the plasmids from live stabs have been midiprepped. The gel showed multiple bands on pSB1A7, also the main band of this one was higher than the others. The rest looked OK. Concentrations of DNA were all between 40 - 100 ng/ul (total volume 500ul) with a 260/280 of 1.8 ± 0.1. | |
| - | + | ||
{{Template:TUDelftiGEM2008_sidebar}} | {{Template:TUDelftiGEM2008_sidebar}} | ||
Latest revision as of 09:24, 27 October 2008
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| July | ||||||
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| August | ||||||
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| 18 | 19 | 20 | 21 | 22 | 23 | 24 |
| 25 | 26 | 27 | 28 | 29 | 30 | 31 |
| September | ||||||
| M | T | W | T | F | S | S |
| [http://2008.igem.org/TUDelft/1_September_2008 1] | [http://2008.igem.org/TUDelft/2_September_2008 2] | [http://2008.igem.org/TUDelft/3_September_2008 3] | [http://2008.igem.org/TUDelft/4_September_2008 4] | [http://2008.igem.org/TUDelft/5_September_2008 5] | [http://2008.igem.org/wiki/index.php?title=TUDelft/6_September_2008&action=edit 6] | [http://2008.igem.org/wiki/index.php?title=TUDelft/7_September_2008&action=edit 7] |
| [http://2008.igem.org/TUDelft/8_September_2008 8] | [http://2008.igem.org/TUDelft/9_September_2008 9] | [http://2008.igem.org/TUDelft/10_September_2008 10] | [http://2008.igem.org/TUDelft/11_September_2008 11] | [http://2008.igem.org/TUDelft/12_September_2008 12] | [http://2008.igem.org/wiki/index.php?title=TUDelft/13_September_2008&action=edit 13] | [http://2008.igem.org/wiki/index.php?title=TUDelft/14_September_2008&action=edit 14] |
| [http://2008.igem.org/TUDelft/15_September_2008 15] | [http://2008.igem.org/TUDelft/16_September_2008 16] | [http://2008.igem.org/TUDelft/17_September_2008 17] | [http://2008.igem.org/TUDelft/18_September_2008 18] | [http://2008.igem.org/TUDelft/19_September_2008 19] | [http://2008.igem.org/wiki/index.php?title=TUDelft/20_September_2008&action=edit 20] | [http://2008.igem.org/wiki/index.php?title=TUDelft/21_September_2008&action=edit 21] |
| [http://2008.igem.org/TUDelft/22_September_2008 22] | [http://2008.igem.org/TUDelft/23_September_2008 23] | [http://2008.igem.org/TUDelft/24_September_2008 24] | [http://2008.igem.org/TUDelft/25_September_2008 25] | [http://2008.igem.org/wiki/index.php?title=TUDelft/26_September_2008&action=edit 26] | [http://2008.igem.org/wiki/index.php?title=TUDelft/27_September_2008&action=edit 27] | [http://2008.igem.org/wiki/index.php?title=TUDelft/28_September_2008&action=edit 28] |
| [http://2008.igem.org/TUDelft/29_September_2008 29] | [http://2008.igem.org/TUDelft/30_September_2008 30] | |||||
| October | ||||||
| M | T | W | T | F | S | S |
| [http://2008.igem.org/TUDelft/1_October_2008 1] | [http://2008.igem.org/TUDelft/2_October_2008 2] | [http://2008.igem.org/TUDelft/3_October_2008 3] | [http://2008.igem.org/wiki/index.php?title=TUDelft/4_October_2008&action=edit 4] | [http://2008.igem.org/wiki/index.php?title=TUDelft/5_October_2008&action=edit 5] | ||
| [http://2008.igem.org/TUDelft/6_October_2008 6] | [http://2008.igem.org/TUDelft/7_October_2008 7] | [http://2008.igem.org/TUDelft/8_October_2008 8] | [http://2008.igem.org/TUDelft/9_October_2008 9] | [http://2008.igem.org/TUDelft/10_October_2008 10] | [http://2008.igem.org/wiki/index.php?title=TUDelft/11_October_2008&action=edit 11] | [http://2008.igem.org/wiki/index.php?title=TUDelft/12_October_2008&action=edit 12] |
| [http://2008.igem.org/TUDelft/13_October_2008 13] | [http://2008.igem.org/TUDelft/14_October_2008 14] | [http://2008.igem.org/TUDelft/15_October_2008 15] | [http://2008.igem.org/TUDelft/16_October_2008 16] | [http://2008.igem.org/TUDelft/17_October_2008 17] | [http://2008.igem.org/wiki/index.php?title=TUDelft/18_October_2008&action=edit 18] | [http://2008.igem.org/wiki/index.php?title=TUDelft/19_October_2008&action=edit 19] |
| [http://2008.igem.org/TUDelft/20_October_2008 20] | [http://2008.igem.org/TUDelft/21_October_2008 21] | [http://2008.igem.org/TUDelft/22_October_2008 22] | [http://2008.igem.org/TUDelft/23_October_2008 23] | [http://2008.igem.org/TUDelft/24_October_2008 24] | [http://2008.igem.org/wiki/index.php?title=TUDelft/25_October_2008&action=edit 25] | [http://2008.igem.org/wiki/index.php?title=TUDelft/26_October_2008&action=edit 26] |
| [http://2008.igem.org/wiki/index.php?title=TUDelft/27_October_2008&action=edit 27] | [http://2008.igem.org/wiki/index.php?title=TUDelft/28_October_2008&action=edit 28] | [http://2008.igem.org/wiki/index.php?title=TUDelft/29_October_2008&action=edit 29] | [http://2008.igem.org/wiki/index.php?title=TUDelft/30_October_2008&action=edit 30] | [http://2008.igem.org/wiki/index.php?title=TUDelft/31_October_2008&action=edit 31] | ||
20th August 2008
Testing the Primers (part 2)
Today another PCR was set up, again using the mastermix and composition also used for the first experiment (18th of August). Only this time the S. cerevisiae genes and cDNA were used. The cDNA contained 2ng/ul and the genes are: ERG8, ERG12, ERG13, HMG2 and MVD1. PCR program (with a 6ºC gradient this time) is as depicted below:
1. 5' @ 95ºC
2. 1' @ 95ºC
3. 1' @ 53.7ºC (gradient of 6C divided over 12 steps)
4. 1' @ 72ºC
5. repeat steps 2-4 29x (total of 30 cycles)
6. 5' @ 72ºC
7. forever @ 4ºC (PCR can be stopped and stored in the fridge at any time from this point on)
This time, the products were put directly on gel. The expected sizes are: ERG8 - ~1400bp, ERG12 - ~1300bp, ERG13 - ~1500bp, MVD1 - ~1200bp and HMG2 - ~3000bp. As there was no visible result on the ERGs gel, only the MVD1/HMG2 gel is shown:
Midiprep
All the plasmids from live stabs have been midiprepped. The gel showed multiple bands on pSB1A7, also the main band of this one was higher than the others. The rest looked OK. Concentrations of DNA were all between 40 - 100 ng/ul (total volume 500ul) with a 260/280 of 1.8 ± 0.1.
