Team:Hawaii/Notebook/2008-10-16

From 2008.igem.org

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(New page: {{Team:Hawaii/Header}} = Things we did today = == Wetlab work == ===Verification of transformants=== :<strong> Grace</strong> [[Image101608colony.png|right|thumb|250px|EtBr stained 2% aga...)
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===Verification of transformants===
===Verification of transformants===
:<strong> Grace</strong>
:<strong> Grace</strong>
-
[[Image101608colony.png|right|thumb|250px|EtBr stained 2% agarose gel ran at 95V for 1 hour.  
+
[[Image:101608colony.png|right|thumb|250px|EtBr stained 2% agarose gel ran at 95V for 1 hour. ]]
{|class=wikitable border=1 align=center
{|class=wikitable border=1 align=center
!Construct
!Construct
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|}
|}
:* Colony PCR of transformants
:* Colony PCR of transformants
 +
::* None of the colonies tested positive for their desired inserts
 +
 +
===Transformation===
 +
:<strong> Margaret</strong>
 +
{|class=wikitable border=1 align=center
 +
!Construct
 +
!Colonies
 +
|-
 +
|align=center|lac promoter/rbs(34)+ rep + pSB1A2
 +
|align=center|0
 +
|-
 +
|align=center|lac promoter/rbs(30)rep/oriV (6) + lac promoter/rbs(34)aada/oriT (A) +pSB1A3
 +
|align=center|113
 +
|-
 +
|align=center|lac promoter/rbs(30)rep/oriV (6) + lac promoter/rbs(34)aada/oriT (A)
 +
|align=center|141
 +
|-
 +
|align=center|lac promoter/rbs(30)rep/oriV (6) + lac promoter/rbs(34)aada/oriT (B) +pSB1A3
 +
|align=center|150
 +
|-
 +
|align=center|oriV (6) + oriT + pSB1A7
 +
|align=center|200
 +
|-
 +
|align=center|pSB1A7 self (did not de-phosphorylate)
 +
|align=center|50
 +
|-
 +
|align=center|DH5-a with RP4
 +
|align=center|3
 +
|-
 +
|align=center|DB3.1 with RP4
 +
|align=center|0
 +
|-
 +
|align=center|(+) pUC18
 +
|align=center|lawn
 +
|-
 +
|align=center|(-) no plasmid
 +
|align=center|0
 +
|}
 +
 +
===Construction of GFP secretion devices===
 +
:<strong>Krystle</strong>
 +
:* Gel purified restricted nir+ rbs and slr+gfpf
 +
:* Dephosphorylated cut pSB1A3 vector
 +
:* Ligation
 +
::* nir+rbs with slr+gfpf
 +
::* lac+rbs with slr+gfpf
 +
:* Transformation into home-made competent and commercially competent DH5α cells and plated on LB amp100 plates
 +
= Discussion =
= Discussion =

Latest revision as of 00:53, 30 October 2008

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

Things we did today

Wetlab work

Verification of transformants

Grace
EtBr stained 2% agarose gel ran at 95V for 1 hour.
Construct Colonies
nir + rbs+GFP+tt in BBpRL1383a-1 2
plac + rbs+GFP+tt in BBpRL1383a-1 0
J33207 in pRL1383a (all plates) lawn
  • Colony PCR of transformants
  • None of the colonies tested positive for their desired inserts

Transformation

Margaret
Construct Colonies
lac promoter/rbs(34)+ rep + pSB1A2 0
lac promoter/rbs(30)rep/oriV (6) + lac promoter/rbs(34)aada/oriT (A) +pSB1A3 113
lac promoter/rbs(30)rep/oriV (6) + lac promoter/rbs(34)aada/oriT (A) 141
lac promoter/rbs(30)rep/oriV (6) + lac promoter/rbs(34)aada/oriT (B) +pSB1A3 150
oriV (6) + oriT + pSB1A7 200
pSB1A7 self (did not de-phosphorylate) 50
DH5-a with RP4 3
DB3.1 with RP4 0
(+) pUC18 lawn
(-) no plasmid 0

Construction of GFP secretion devices

Krystle
  • Gel purified restricted nir+ rbs and slr+gfpf
  • Dephosphorylated cut pSB1A3 vector
  • Ligation
  • nir+rbs with slr+gfpf
  • lac+rbs with slr+gfpf
  • Transformation into home-made competent and commercially competent DH5α cells and plated on LB amp100 plates

Discussion

Quote of the Day

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


[http://manoa.hawaii.edu/ Sponsor_UHM.gif][http://manoa.hawaii.edu/ovcrge/ Sponsor_OVCRGE.gif][http://www.ctahr.hawaii.edu Sponsor_CTAHR.gif]


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