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.
- 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
- 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
- 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.