"
Template:Team:UC Berkeley/Notebook/SC notes
From 2008.igem.org
| Line 1: | Line 1: | ||
== 2 July 2008 (W) == | == 2 July 2008 (W) == | ||
Today was a busier day. | Today was a busier day. | ||
| + | |||
A.M.: Finished construction files from yesterday. | A.M.: Finished construction files from yesterday. | ||
Revision as of 00:52, 3 July 2008
Contents |
2 July 2008 (W)
Today was a busier day.
A.M.: Finished construction files from yesterday.
P.M.: We started the assembly work today! After picking out 24 basic parts with C/A vectors and 24 with K/A vectors, I helped out with digestion; did purification; and watched ligation.
1 July 2008 (T)
Today: We continued to help with electronically sorting the basic parts later to be used in composite part building. In the earlier part of the afternoon, we picked colonies for the vectors that were being tested -- since we were desperately out of things to do. Towards evening, we were given the task of creating oligos and making construction files for a couple variations on existing basic parts. I was responsible for seven construction files: {rbs_barstar>}; {prepro>}, {a~prepro>}, and {rbs_prepro>} of both PhoA and LamB proteins.
Monday: There was not much for us to do during the morning. The vectors were still being tested to see whether they were functional, before we scale up and start doing actual assembly of composite parts. In the afternoon, however, we helped with electronically sorting out basic parts (with the C/A and A/C vectors) from the main list of basic parts. As for actual lab action: I learned to do the Agencourt SPRI cleanup on a couple of the vectors that were being tested.
28 June 2008 (Sa)
I wasn't working in the lab today; but, my sequencing results came back! All four of my tags have perfect reads.
27 June 2008 (F)
Today, we've completed one round of lab work from beginning to end -- from researching literature to sending out samples for sequencing! We finished doing minipreps in the morning, and (literally) taped our samples to the wall outside to be sent for sequencing. The results should come by tomorrow (Sat), so that we can analyze them! We also learned to run an analytical gel (using E-gel mode) on Molly's colony PCR to see if the products matched with the predicted size. At the end of the day, the team started planning out duties for everyone over the next couple weeks when we start building composite parts. Everything would be on a much larger scale; so the tactic is assign each person one specific job in the assembly line of jobs. As of present, I'll be handling the purification process (after digestion and before ligation).
26 June 2008 (Th)
It was a much lighter day today. I picked out two colonies to grow overnight tonight, and will miniprep them tomorrow. Since we had extra time on our hands, we also did a couple minipreps for Madhvi.
25 June 2008 (W)
Our busy schedule today:
We purified the PCR product (using Zymo cleanup); set up for restriction digest; cleaned up (Zymo) the digest; set up for Digestion; did Transformation; and, we learned how to use the "spreader method" to plate colonies -- without setting the ethanol on fire...
We finished at 8:15p.m.
24 June 2008 (T)
I finished creating/organizing my oligos and construction files onto spreadsheets and documents.
We did set up for the wobble PCRs; they're in the thermocycler overnight until tomorrow!
23 June 2008 (M)
Having completed one week of training/lectures, I've finally started on my minute portion of the lab work! Over last weekend, I searched for the DNA sequences of the FLAG-tag and the AP-tag. The objective was to find an existing E. coli vector for said sequences. Upon completing the search, I designed oligos for EIPCR for small parts under 30 bp (like our tags). When we discovered, however, that our sequences were too long - leaning precariously over the "30bp border" - we decided to prepare for a Wobble (overlap extension) PCR instead. This involved designing Fwd/Rev oligos that had a 20bp overlap. When that was done, we ordered the oligos. We now eagerly expect their arrival tomorrow at around 3p.m., when we can actually do the PCR reation!
