Team:Hawaii/Notebook/2008-08-23

From 2008.igem.org

(Difference between revisions)
(3A assembly)
(Verification of transformants (cont.))
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:* Plasmid prep of nir+rbs, rbs+GFP
:* Plasmid prep of nir+rbs, rbs+GFP
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:* Overnight RE digest of plasmid preps, nir plasmid, GFP plasmid with XbaI and SpeI
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{|class=wikitable border=1 align=center
 +
!Plasmid prep
 +
!DNA concentration
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!260/280
 +
!260/230
 +
|-
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|nir+rbs
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| 394.63 ng/μl
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|2.04
 +
|2.36
 +
|-
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|rbs+GFP
 +
|94.9 ng/μl
 +
|2.06
 +
|2.17
 +
|}
===3A assembly===
===3A assembly===

Revision as of 01:40, 24 August 2008

Projects Events Resources
Sponsors Experiments Milestones Protocols
Notebook (t) Meetings (t)

Things we did today

Wetlab work

Verification of transformants (cont.)

Grace
  • Plasmid prep of nir+rbs, rbs+GFP
Plasmid prep DNA concentration 260/280 260/230
nir+rbs 394.63 ng/μl 2.04 2.36
rbs+GFP 94.9 ng/μl 2.06 2.17

3A assembly

Grace
File:082308REdigests.jpg
EtBr stained 2% agarose gel ran at 60V for 3 hours. Thirty microliters of RE digest reaction were loaded into each well.
  • Gel purified RE digests
  • Ligated with pSB1A2:
  • nir+rbs
  • plac+rbs
  • rbs+GFP
  • GFP+tt
  • GFPf+tt
  • Transformed DB3.1 cells with 5μl ligation reaction

BB-pRL1383a MCS re-replacement

Grace
  • Gel purified RE digests
  • Ligated J33207 with BB-pRL1383a
  • Transformed DH5&alpha cells with 5 μl ligation reaction

Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


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