IGEM:IMPERIAL/2006/LabCalendar/2006-8-18: Difference between revisions
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*2H | *2H | ||
**Unsuccessful, needs redoing | **Unsuccessful, needs redoing | ||
* | **Second miniprep unsuccessful. Have to let MIT know we have been struggling to ligate part 2H and that we suspect that it may involve a mutation in the Spe1 site. | ||
==Ligations== | ==Ligations== | ||
Revision as of 09:25, 21 August 2006
Maxipreps
- 12D - for J37020 ligation
- 5M - for ligating into aiiA construct when primers arrive
- J37016 - so a sample can be sent for sequencing
Sequencing
- J37016 sent for sequencing
- 1 μg DNA
- 12.5 pmol I13504 primer (CAA CTA GCA GAC CAT TAT C)
Minipreps
- J37015RS
- We have innoculated a 100 mL culture in the fridge to be maxipreped on Monday
- All cultures were successful; however, we took only colony 4 since it showed the best looking bands on the gel (yes, I guess we do have to be discriminatory)
- 1M
- Band at around 2000 showing the plasmid, the part itself is too small and may have run too far, so we assume that it is correct
- This part can be maxipreped on Monday along with J37015RS
- 4G
- Successful miniprep, we can start testing this on Monday as it is an entire part ready to be testing. Please ensure that you culture in Kanamycin LB (located in fridge)
- 2H
- Unsuccessful, needs redoing
- Second miniprep unsuccessful. Have to let MIT know we have been struggling to ligate part 2H and that we suspect that it may involve a mutation in the Spe1 site.
Ligations
- J37031 = J37019 + B0015 (1I)
- J37032 = R0062 (9G) + I13504 (12D)
- 12D on the gel after digestion was not clean, this might pose a small problem, but I cut out the smaller band anyway that look around 800 bp (compared to the J37019 which is also about 800 bp).
- Electroporations this afternoon
