IGEM:IMPERIAL/2006/LabCalendar/2006-9-4
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TO DO: *Testing of T9002, J37016, J37020, S01656, J37015, J37015RS *Culture J37036 & J37018 from plate in incubator for maxi & mini on Tuesday *Make frozen stock of J37022 & culture up for testing ! *Send J37024 for sequencing !!! *Order new maxiprep kit !!! *Need more autoclaved flasks !!! *Need more isopropanol too!
ligations
- J04500 (vector) and J37024 (insert)being ligated overnight
LoxP PCR
- Was an amazing success
- Jonny is a hero
LoxP PCR fusion
Western Blot
Maxiprep
- Maxiprep new AiiA gene from overnight culture.
- Not very much bacteria (hence DNA) present.
AiiA PCR
- Using the new primers and the newly arrived AiiA gene (maxi-preped this morning), PCR carried oput to add the LVA tag and immunotag.
- PCR annealing temperatures calculated to be 10 cyles at 51[[:Category:{{{1}}}|{{{1}}}]] and 20 cycles at 69[[:Category:{{{1}}}|{{{1}}}]]
Culturing
- S01656 +IPTG, S01656 -IPTG, J37016 was cultured for testing
- J37036 & J37018 & T9002 could not be cultured since no colonies could be seen on the plate - they have to grow overnight
Frozen Cells
- Made a frozen stock of cells: J37025 (3 different colonies, but all of them have been verified to be correct)
