Plates to check
- K235003, 1 and 4 colonies
- K235005, 1 and 0
- K235010, 4 and 6
- J04450-Kan 7A, 21 and 24
- J04450-Kan 7G, 3 and 2
- P1010 Kan, many colonies
- Amp, many
- Cm, none
- I0500, quite a few
- R0051, many
So the stab of P1010 Cm is the only one not to grow at all and we have it already anyway.
12.30pm
2 broth cultures started of everything that grew. Cultures not left shaking at this point.
4.00pm
- Add IPTG to one of I0500, one of J04450 7A, and one of 7G to a concentration 0.83mM
- Start shaking before we leave
5:20pm
Kyliah added a number of parts to the assemblies page, and added most of those parts to the actual iGEM team page (!) woohoo...
It looks like, to finish the NAND gate proper, we have four rounds of assemblies to do, each set dependent upon the last:
- K235009, K235015, K235016, K235017
- K235018, K235019, K235020, K235021
- K235022, K235023, K235024, K235011
- K235025, K235026, K235027, K235028
K235011 makes an mCherry analogue to K145303 - testing of ribolock temperature sensitivity?
K235023 makes an interesting test signal inverter
The last batch makes the NAND gate proper with three positive controls
Each round will take at LEAST two days (one to miniprep, digest and ligate, and transform and grow overnight; one to broth culture overnight). So, in theory, by the middle of September, Natasha and Kyliah can have everything assembled and ready for final testing (with some minor tests along the way to fill time on broth culture days).
We still need to look into having some of the submittable parts sequenced... Which ones should we do? And should we wait to send them in a batch?
Comments (2)
Chris Tuttle said
at 7:59 pm on Sep 3, 2009
One big thing I don't undestand from your last few days work is where are all the gels? do we have confirmation of part length (even approximate) for any of the parts you say are finished? how close are we to putting together the simple test system that I had described to natasha so that we can have even a rough estimate of what % of colonies contain properly assembled parts? Also I thought we were going to pick 7 colonies from each part.
Natasha Tuskovich said
at 10:49 am on Sep 4, 2009
We will be able to run gels today, now that we have some new working parts. The test system had one part go wrong and that will be retried today or next week. We went with two colonies for each part because we still ended up with 17 cultures and we ran out of glassware and holder space with that many.
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