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Team:The University of Alberta/26 May 2008
From 2008.igem.org
NK 130-530
Hey guys, Kelly and I (Anthony) were in on Friday and we managed to excise the bands and do the gel purification. Kelly has put Winnie's digested producs in the -20, I have put the purification products also in the -20 (wrapped together in green tape and labelled). As well, anything that was left out on the bench was put in the -20 because we have no clue what was in those tubes. Kelly and I were having a difficult time following the lab book, so PLEASE be explicitly clear about what has been performed, what is in each of the biobricks, etc... It might seem annoying and redundant but the volunteers have simply no way of following otherwise. A simple summary of each procedure and protocol would be GREATLY appreciated! We didn't have time to do to the sequencing reaction because Kelly has left for Rocky Mountain, and I am not about to start a 2.5 hr reaction at 7:30 PM on a Friday night!
Good luck with things on Monday, and I will see you Tuesday
Anthony
Contents |
Today In The Lab
- Overday's of Purple Russian and Tryp
- Finish Fundraising Letter
- Finish Construction of the Binary Vector
- Plant 2 more trays of arabidopsis
- Work with the Butanol Pathway some more
Jason:
- I have made overday's of Blue Ox as well as Purple Russian and Tryp.
- Finished writing the first draft of the Fundraising letter
Chris:
- Got the sequences from last week (21, 22, 35, and 99). I tried BLASTing them but I didnt get a single significant match for either part (the only result was a poor match to some expression vector). I tried to do a [http://blast.ncbi.nlm.nih.gov/bl2seq/wblast2.cgi| Blast 2] on each part to the sequences from last year, but the results were not good:
- 21: The sequence matched well with the sequence from last year. Did a normal BLAST on last years sequence and it worked fine; makes me wonder why I had no results when blasting this year's sequence.
- 22: There was no significant match to last year's sequence at all aside from a 41bp match at the start of the sequences.
- 25: I couldnt do a BLast2 on this part because there was no reference sequence from last year. This part (Butanol dehydrogenase) did have a sequence but it was as a part of a larger BioBrick with another part (butyraldehyde dehydrogenase). Trying to Blast2 these sequences resulted in to significant matches.
- 99: There was no reference sequence from last year for this part.
Part 23 was not sequenced (did we forget?). Should do this before we do anything with it to make sure its what we think it is! EDIT: Turns out that the parts were mislabelled....What we labelled as 22 is actually part 23 (and 23 is likely to be part 22). This means we have the sequence for the real 23....but we have no sequence for part 22.
Tom:- Summarized our near future butanol plans on page 37 of the masterbook.
- Digested I0500 with Spe/Pst and R3298 (butanol DH - I725025) with Xba/Pst.
- The reason for redoing digestion of I725025 is because last week's band did not match our prediction.
- Ran the colony PCRs and the two digests above on a gel (similar to the gel ran on May 23 since no picture was taken from that gel).
- Did NOT run 21 however; could not find the PCR colony for those 4 tubes.
- Gel excision and ligation of I0500.
- Won't be able to transform the ligations as we are to incubate at room temperature for 3 hours.
- If you could would please make Glycerol stocks as well as a mini prep on those Overdays that Jason has done
Volunteers Scheduled for Today
5-8 AS
5-8 KR
5-8 SG
