IGEM:Harvard/2007/Laboratory Notebooks/Quorum Sensing/Week 3

07/02/07

Sequencing Parts

Stephanie Nanodropped and sent the following parts for sequencing:

• S03623
• S03608
• J37034
• J23039
• R0011
• R0140
• T9002
• B0015
• R0051

Stephanie's notebook entry

Constructing Growth Curves

Stephanie used the plate reader and Bl21+I13263 cells diluted 1:10, 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, and 1:100 as well as to construct a growth curve of OD vs time.

Stephanie's notebook entry

Liquid Cultures

Stephanie grew liquid cultures of I5311, OmpA1+his, I13522, T9002, and I13263 in preparation for induction on 07/03/07.

Stephanie's notebook entry

Digestion

Perry performed the following digests:

• R0011 with SpeI and PstI
• R0040 with SpeI and PstI
• R0051 with SpeI and PstI
• S03608 with EcoRI and SpeI
• S03623 EcoRI and SpeI
• J37034 XbaI and PstI (x2)
• B0015 EcoRI and SpeI

Perry and Stephanie also ran a gel and gel-extracted the result. However, the J37034 did not turn out as expected so Stephanie ran another digest on the J37034

Perry's notebook entry

07/03/07

Results of the Growth Curves

Results posted by Stephanie

Digests

Perry and Stephanie digested the E0240, S03608, S03623, and B0015 parts in order to rebuild the J37034.

Stephanie's notebook entry Perry's notebook entry

Ligation and Transformation of Parts

Perry and Stephanie ligated with Quick Ligase: 7ul S03608/S03623 + 3ul B0015 in pSB1AK3. They then transformed the ligated parts into 20ul Top10.

Perry's notebook entry