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Team:TU-Munich/Notebook/Labjournal
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* CaCl2 competent ''E. coli'' XL1-Blue cells were put out from the stock in -80 °C freezer and were gently thawed on ice. | * CaCl2 competent ''E. coli'' XL1-Blue cells were put out from the stock in -80 °C freezer and were gently thawed on ice. | ||
| - | * 2 µl of DNA was added to 100 µl of competent cells and gently mixed. | + | * 2 µl of DNA was added to 100 µl of competent cells and gently mixed. |
* 30 min incubation on ice | * 30 min incubation on ice | ||
| - | * 5 min. heat shock at | + | * 5 min. heat shock at 37 °C |
* Adding of 1ml LB-medium to each tube. | * Adding of 1ml LB-medium to each tube. | ||
| - | * Incubation for | + | * Incubation for 45 min at 37 °C in the 180 rpm cell-culture shaker. |
| - | * | + | * 100 µl of the cell suspension was plated on one chloramphenicol plate. |
| - | * The rest were centrifuged for 1 | + | * The rest were centrifuged for 1 min at 13000 rpm and the supernatant was dicarded. |
| - | * The pellet was resuspended in | + | * The pellet was resuspended in 100 µl of LB-medium and this concentrated cell suspension was plated again on a new chlorampenicol plate. |
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Labjournal
Week 1
Monday, April 22nd
Transformation of E.coli XL1 blue with Phytochrome B (2-908 N-terminal amino acids) (BBa_K801031, RFC25, pSB1C3)
Investigator: Jeff, Leonie, Rosario
Aim of the experiment: Transformation of Phytochrome B for protein fusion.
Procedure:
- CaCl2 competent E. coli XL1-Blue cells were put out from the stock in -80 °C freezer and were gently thawed on ice.
- 2 µl of DNA was added to 100 µl of competent cells and gently mixed.
- 30 min incubation on ice
- 5 min. heat shock at 37 °C
- Adding of 1ml LB-medium to each tube.
- Incubation for 45 min at 37 °C in the 180 rpm cell-culture shaker.
- 100 µl of the cell suspension was plated on one chloramphenicol plate.
- The rest were centrifuged for 1 min at 13000 rpm and the supernatant was dicarded.
- The pellet was resuspended in 100 µl of LB-medium and this concentrated cell suspension was plated again on a new chlorampenicol plate.
