IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/08/24

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Working on LovTAP and LuxY synthesized plasmids from Mr.Gene: Edinburgh Shipment
I am going to do PCR reactions in order to amplify the LovTAP and LuxY construction synthesized. These products will be send to Edinburgh team.

The products obtained will be purified using the High Pure PCR Product Purification kit from Roche.

Primers:

Forward:Preffix primer

Reverse:Suffix primer

PCR template:

Plasmid harboring LovTAP and luxY from Mr.Gene


 * Reaction 1


 * Reaction 2


 * 35 PCR programmed cycles:

The reaction starts at 95°C during 5 min. After this step, the reaction 2 is loaded to the reaction 1 sample.

Each cycle is programmed as follows:

After the cycle 35, the temperature changes to 72°C during 10 min and ends at 4°C.

Results:PCR reactions LovTAP and LuxY
Four PCR reactions were done for LovTAP and LuxY. The resultant amplified products are showed in the next image. From lane 8 to lane 11, the products are of the LuxY PCR reactions, the strongest band correspond to LuxY with size 820nt. As there were other products, I am going to extract LuxY from gel, as was done previously  for LovTAP.

In LovTAP PCR reactions, the amplified products were around the expected size -889 nt- that is the lenght of LovTAP synthesized. These PCR reactions will be purified using the High Pure PCR Product Purification kit from Roche.




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