EPF-Lausanne/9 July 2009

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(New page: ===Wet Lab=== 1. Miniprep and isolations of the yesterday transformed plasmids. (cf. [https://2009.igem.org/Team:EPF-Lausanne/Notebook/Wet_Lab#08.07.09 08.09.09 subpart]) Concentrations of...)
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Revision as of 15:40, 27 July 2009

Wet Lab

1. Miniprep and isolations of the yesterday transformed plasmids. (cf. 08.09.09 subpart)

Concentrations of the plasmids: cf. lab notebook pp. 8-9


2. PCR of pCal-n to PCR up LOVTAP with the different primers designed the 06.09.09, the products are:
- Prom_T7-RBS-CBP-LOVTAP
- RBS-CBP-LOVTAP
- CBP-LOVTAP
- LOVTAP

Result: Prom_T7-RBS-CBP-LOVTAP didn't worked, the other were ok.

3. An agarose gel was runned to check PCR products


4. PCR products were digested with EcorI and SpeI and [http://partsregistry.org/Part:BBa_B0010 BBa_B0010] (plasmid chosen containing the terminator) was digested with EcorI and XbaI, the digestion products were treated with phosphatase. Then, PCR products were purified.

Finally, LOVTAP (PCR products) were ligated on [http://partsregistry.org/Part:BBa_B0010 BBa_B0010] (plasmid chosen containing the terminator).


5. Two more iGEM parts have been transformed:

Parts&Characteristics
Part Characteristic Resistance Well (Kit Plate)

[http://partsregistry.org/Part:BBa_I6007 BBa_I6007]

Double repressor: called Inverter TetR

A

1C (2)

[http://partsregistry.org/Part:BBa_P1010 BBa_P1010]

Death Cassette

C

5E (1)


Remarks: [http://partsregistry.org/Part:BBa_P1010 BBa_P1010], the death gene has to be transformed in ccdB (death gene) resistant cells: One Shot ccdB survival 2T1 E. Coli