IGEM:IMPERIAL/2006/LabCalendar/2006-8-11

Control Digests for finalized prey cell J37015 & pSB1A2

 * Gel of the digests from yesterday was run and assessed: See results on the right.
 * Gel: (from left to right)
 * Lanes 1-3: Recombinant Plasmid A (Digest 1-3)
 * Lanes 4-6: Insert Plasmid 12D->24A (Digest1-3)
 * Lanes 7-9: Vector Plasmid 6B (Digest 1-3)
 * Lane 1, 2, 3, 4, 7, 8, 9 have bands at expected lengths
 * Other unexpected lengths might have occured due to contamination - and probably PvuII did not work correctly.
 * Overall, suggests that the sequence of the plasmid is correct.

Culture

 * J37015RS and J37016 being cultured in liquid media for miniprep this afternoon
 * 2 plates of J37015RS and 3 plates of J37016 are being analysed, each containing 6 colonies each

PCR

 * Dr. Mann's PCR was successful for the immunotag, however was unsuccessful for the LoxP sites
 * This is being redone using slightly different temperatures
 * The Cre part was successfully PCR-ed without the help of Dr.Mann
 * Four Cre parts was cut out of the gel available for ligating

Ligations

 * AiiA was given to us by Dr. Mann in gel format and we isolated the Cre parts (above)
 * These were ligated into the hanging T plasmid Dr. Mann gave us
 * Ligation started at 14:30

To do tomorrow

 * Gene Clean Ligations Cre & AiiA parts
 * Maxiprep 2H again
 * Miniprep J37015RS and J37016

NO PCR on next Tuesday only one of the machines. Will be returned on wednesday. Will be used again on wednesday or thursday.