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Team:Calgary/6 July 2010
From 2010.igem.org
(Difference between revisions)
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Himika: | Himika: | ||
* Today I reran the transformation that I did yesterday July 5th where I constructed E0040 and pSB1AC3 and plate it on chloramphenicol. | * Today I reran the transformation that I did yesterday July 5th where I constructed E0040 and pSB1AC3 and plate it on chloramphenicol. | ||
| - | * I also miniprepped the overnight cultures from last night. I10504 and | + | * I also miniprepped the overnight cultures from last night. I10504 and I13507. The only cultures that grew were I10504 and I miniprepped 6 colonies from 1 plate. |
* I also constructed E1010 with B0015. | * I also constructed E1010 with B0015. | ||
| Line 12: | Line 12: | ||
* Procedure: | * Procedure: | ||
** ''Insert'' | ** ''Insert'' | ||
| - | ** E1010 (pSB2k3) cut with EcoI/SpeI | + | ** E1010 (pSB2k3) cut with EcoI/SpeI 5uL DNA |
| - | ** Buffer I was used 3. | + | ** Buffer I was used 3.5uL |
| - | ** 0.5 | + | ** 0.5 uL of EcoRI and 0.5 uL of SpeI |
** 25.5 uL of water | ** 25.5 uL of water | ||
** ''Vector'' | ** ''Vector'' | ||
| - | ** B0015 (pSB1AK3) cut with EcoI/XbaI | + | ** B0015 (pSB1AK3) cut with EcoI/XbaI 5uL DNA |
| - | ** Buffer I was used 3. | + | ** Buffer I was used 3.5uL |
| - | ** 0.5 | + | ** 0.5 uL of EcoRI and 0.5 uL of XbaI |
** 25.5 uL of water | ** 25.5 uL of water | ||
| - | * The vector was phosphatased with | + | * The vector was phosphatased with 5uL water, 4uL Buffer and 1uL phosphatase. The solution was left in 37 degree Celcius for 30 minutes and heat killed for 10 minutes in 65 degree Celcius. |
* 5ul of the vector and the insert in a tube and add 10 ul of quick ligase buffer and 1 ul of quick ligase. The solution was left in room temperature for 10 minutes and transformed. | * 5ul of the vector and the insert in a tube and add 10 ul of quick ligase buffer and 1 ul of quick ligase. The solution was left in room temperature for 10 minutes and transformed. | ||
Revision as of 22:19, 6 July 2010
Tuesday July 6, 2010
Happy Birthday Paul!!
Himika:
- Today I reran the transformation that I did yesterday July 5th where I constructed E0040 and pSB1AC3 and plate it on chloramphenicol.
- I also miniprepped the overnight cultures from last night. I10504 and I13507. The only cultures that grew were I10504 and I miniprepped 6 colonies from 1 plate.
- I also constructed E1010 with B0015.
- Procedure:
- Insert
- E1010 (pSB2k3) cut with EcoI/SpeI 5uL DNA
- Buffer I was used 3.5uL
- 0.5 uL of EcoRI and 0.5 uL of SpeI
- 25.5 uL of water
- Vector
- B0015 (pSB1AK3) cut with EcoI/XbaI 5uL DNA
- Buffer I was used 3.5uL
- 0.5 uL of EcoRI and 0.5 uL of XbaI
- 25.5 uL of water
- The vector was phosphatased with 5uL water, 4uL Buffer and 1uL phosphatase. The solution was left in 37 degree Celcius for 30 minutes and heat killed for 10 minutes in 65 degree Celcius.
- 5ul of the vector and the insert in a tube and add 10 ul of quick ligase buffer and 1 ul of quick ligase. The solution was left in room temperature for 10 minutes and transformed.
- I also did research on the ibpA/ ibpB pathways in order to present on to Dr. Schryvers on Friday.
- I also overnight cultured I10507 and plated the contstruct.
