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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) to see if a certain set of PCR products matched with the predicted size.

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!



Sherine Cheung
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