Team:Hawaii/Notebook/2008-09-25

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= Things we did today =
== Wetlab work ==
===Verification of Ligation===
:<strong> Margaret </strong>

:* The plac/rbs+rep and oriTr(BB) transformations resulted in 0 colonies with the correct insert, so the remainder of the ligation reaction from 9/16 was digested with XbaI and run on a gel. 

===Sequencing & Colony PCR===
:<strong> Margaret </strong>

:*PCR
::* oriV colonies (3,4,6,8,9,10,11)
::*aadA(BB) colonies (4,5,7,8)
::*aadA(pRL) colonies (1,2)
::*oriT + J33207 (1,2,4)
::*oriT + J33207(de-phosphorylated) colony 1
::*5 additional colonies of oriTr transformation were PCR'ed


===Construction of secretion device and redo of BB-pRL1383a (cont.)===
:<strong> Grace</strong>
[[Image:092508REdigest.jpg|right|thumb|300px|EtBr stained 2% agarose gels ran at 60V for 1.5 hours and 2 hours, respectively. Thirty microliters of RE digested products were loaded into each well.]]
:* Ran RE digests from last night on gel
::* nrsg8 and nrg4 did not cut. Implies insert wrong?
:* Extracted bands from gel
:* Ligated:
::* 3A using pSB1A3:
:::* nrg7 + tt
:::* rgt1 + nir
:::* rgt1 + plac
:::* rgt2 + nir
:::* rgt2 + plac
:::* pSB1A3 (control)
:* pRL1383a + J33207
::* pRL1383a to self (control)
:* Transformed into DH5&alpha; using 5 &mu;l ligation reaction; also transformed J33207 and pRL1383a plasmids as control)

= Discussion =

= Quote of the Day =
<blockquote>''History is the only laboratory we have in which to test the consequences of thought.'' - Étienne Gilson</blockquote>

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