IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/09/02

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Working on cI inverter construction, pSB3K3 backbone and E0240 part
Once the plasmids harboring the parts showed in the next table were correctly isolated,  I am going to digest some with EcoRI and PstI and the others with XbaI and PstI in order to fuse them with their corresponding part.

K098991 preparation for ligation with plasmid pSB3K3 and J04450 preparation for ligation with promoters-LovTAP, trpL-cI inverter and trpL-GFP constructions for characterization.'''
 * '''Restriction enzymes EcoRI and PstI:

Plasmids used:

Plasmid harboring K098991 isolated from colony 8.

Plasmid harboring J04450 isolated from colony 8. Two replicas

P0451 and E0240 preparation for ligation with constitutive promoter (J23101) and trpL promoter '''
 * '''Restriction enzymes XbaI and PstI:

Plasmids used:

Plasmid harboring P0451 isolated from colony 6.

Plasmid harboring E0240 isolated from colony 5. Two replicas

The reactions were incubated at 37°C overnight.

Note: Inactivate the enzymes at 80°C during 10 min.

Results restriction enzyme assays


The sizes of the digested products are around the expected lenght for each part.


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