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TUDelft/22 August 2008
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{{Template:TUDelftiGEM2008_calendar}} | {{Template:TUDelftiGEM2008_calendar}} | ||
| - | = | + | =August 22nd= |
==DNA extraction test== | ==DNA extraction test== | ||
| - | Today another gel was run with DNA soaked from the spots. To test the soaking-protocol, various steps of the protocol were performed on the DNA. This time 10 ul of TE was used due to the soaking of the water | + | Today another gel was run with DNA soaked from the spots. To test the soaking-protocol, various steps of the protocol were performed on the DNA. This time 10 ul of TE was used, because due to the soaking of the water into the punch, using 5 ul reduced free liquid to be added to cells (or for analysis) below 2 ul. Again various conditions were tested, and also the later sent spots from the promoter strength testing DNA were analyzed. To give an idea of DNA concentrations, all spots were nanodropped. These results are in the table below. For this testing we used a spot we won't be using and showed a good band on the gel of the repository. 1002 9A, a lock from Berkeley06, which had inconsistent sequence. |
{| border="1" | {| border="1" | ||
| Line 11: | Line 11: | ||
! Sample !! ng/ul !! A260/A280 | ! Sample !! ng/ul !! A260/A280 | ||
|- | |- | ||
| - | |TE ( | + | |TE (blank) |
|0.53 | |0.53 | ||
| 1.42 | | 1.42 | ||
| Line 23: | Line 23: | ||
|0.89 | |0.89 | ||
|- | |- | ||
| - | |10 ul TE + 1002 9A punch, T = | + | |10 ul TE + 1002 9A punch, T = 42ºC |
|11.04 | |11.04 | ||
|0.75 | |0.75 | ||
|- | |- | ||
| - | |10 ul TE + 1002 9A punch, T = | + | |10 ul TE + 1002 9A punch, T = 42ºC + Spinned |
|11.78 | |11.78 | ||
|1.23 | |1.23 | ||
| Line 44: | Line 44: | ||
|} | |} | ||
| + | ''Tr = Room Temperature'' | ||
| + | |||
These results have again rather low 260/280, but DNA concentrations that should be visible on a gel. Concentrations this time with increased following of iGEM procedure. | These results have again rather low 260/280, but DNA concentrations that should be visible on a gel. Concentrations this time with increased following of iGEM procedure. | ||
Latest revision as of 08:52, 27 October 2008
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| July | ||||||
| M | T | W | T | F | S | S |
| 1 | 2 | 3 | 4 | 5 | 6 | |
| 7 | 8 | 9 | 10 | 11 | 12 | 13 |
| 14 | 15 | 16 | 17 | 18 | 19 | 20 |
| 21 | 22 | 23 | 24 | 25 | 26 | 27 |
| 28 | 29 | 30 | 31 | |||
| August | ||||||
| M | T | W | T | F | S | S |
| 1 | 2 | 3 | ||||
| 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| 11 | 12 | 13 | 14 | 15 | 16 | 17 |
| 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] | ||
August 22nd
DNA extraction test
Today another gel was run with DNA soaked from the spots. To test the soaking-protocol, various steps of the protocol were performed on the DNA. This time 10 ul of TE was used, because due to the soaking of the water into the punch, using 5 ul reduced free liquid to be added to cells (or for analysis) below 2 ul. Again various conditions were tested, and also the later sent spots from the promoter strength testing DNA were analyzed. To give an idea of DNA concentrations, all spots were nanodropped. These results are in the table below. For this testing we used a spot we won't be using and showed a good band on the gel of the repository. 1002 9A, a lock from Berkeley06, which had inconsistent sequence.
| Sample | ng/ul | A260/A280 |
|---|---|---|
| TE (blank) | 0.53 | 1.42 |
| 10 ul TE + 1002 9A punch, Tr | 4.94 | 0.69 |
| 10 ul TE + 1002 9A punch, Tr + spinned | 9.69 | 0.89 |
| 10 ul TE + 1002 9A punch, T = 42ºC | 11.04 | 0.75 |
| 10 ul TE + 1002 9A punch, T = 42ºC + Spinned | 11.78 | 1.23 |
| 10 ul TE + promoter strength punch, Tr | 5.94 | 0.97 |
| 10 ul TE + promoter strength punch, 42 C + spinned | 4.81 | 1.36 |
| 10 ul TE + 3 ul pSB1A7 (isolated on 20-08-08) | 11.69 | 1.66 |
Tr = Room Temperature
These results have again rather low 260/280, but DNA concentrations that should be visible on a gel. Concentrations this time with increased following of iGEM procedure.
The gel showed no visible DNA bands for the soaked punches. The positive control did show bands, as can be seen below.
Transformation
A transformation has been started today for testing of the midiprepped vectors. To prepare for the arrival of our thermosensitive sequences next week we also tried to transform a couple of parts for our constructs, although if these fail we will need a different way to obtain the parts. The parts (apart from the midiprepped) used for a transformation are:
| Part | ng/ul | A260/A280 |
|---|---|---|
| I712019 | 14.22 | 0.64 |
| B0015 | 14.48 | 0.68 |
| I13453 | 10.16 | 0.88 |
| R0010 | 4.44 | 0.66 |
| B0030 | 11.33 | 0.67 |
| I742152 | 20.2 | 0.69 |
Punches were no longer obtained by the punch tool but by cutting them out with a scalpel, due to time gain and the punch tool not working very well. All these punches were soaked in 10 ul TE and transformations were started with 5 ul of extracted DNA. After transformation cells were plated on LB+Amp.
