Team:Hawaii/Notebook/2008-10-15

From 2008.igem.org

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(Drylab Work)
(Construction of secretion device (cont.))
 
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:::* Plated 25 μl, 50 μl, 75 μl, and 100 μl plates in hopes of isolating single colonies
:::* Plated 25 μl, 50 μl, 75 μl, and 100 μl plates in hopes of isolating single colonies
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:<strong> Krystle </strong>
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:* PCR
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::* nir+rbs
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::* slr+gfpf
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:*Restriction digest overnight
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::* nir+rbs with EcoRI and SpeI
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::* slr+gfpf with XbaI and PstI
 +
::* pSB1A3 with EcoRI and SpeI
 +
 +
=== Ligation===
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:<strong> Margaret</strong>
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 +
:* lac promoter/rbs(34) (E,S) + rep2 (X,P) + pSB1A3 (de-phosphorylated, E,P)
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:* lac promoter/rbs(30)rep/oriV(6) (E,S) + lac promoter/rbs(34)aada/oriT(A) (X,P) +pSB1A3 (de-phosphorylated, E,P)
 +
:* lac promoter/rbs(30)rep/oriV(6) + lac promoter/rbs(34)aada/oriT (A)
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:* lac promoter/rbs(30)rep/oriV (6) (E,S) + lac promoter/rbs(34)aada/oriT(B) (X,P) +pSB1A3(de-phosphorylated, E,P)
 +
:* oriV (E,S)  + oriT (X,P) + pSB1A7 (E,P)
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:* pSB1A7 self (did not de-phosphorylate) (E,P)

Latest revision as of 00:53, 30 October 2008

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Things we did today

Wetlab work

Construction of secretion device (cont.)

Grace
  • Transformed DH5α
  • Used 5 μl of n/p + rgt ligation reaction
  • Used 1 μl of J33207/pRL1383a ligation reaction
  • Plated 25 μl, 50 μl, 75 μl, and 100 μl plates in hopes of isolating single colonies
Krystle
  • PCR
  • nir+rbs
  • slr+gfpf
  • Restriction digest overnight
  • nir+rbs with EcoRI and SpeI
  • slr+gfpf with XbaI and PstI
  • pSB1A3 with EcoRI and SpeI

Ligation

Margaret
  • lac promoter/rbs(34) (E,S) + rep2 (X,P) + pSB1A3 (de-phosphorylated, E,P)
  • lac promoter/rbs(30)rep/oriV(6) (E,S) + lac promoter/rbs(34)aada/oriT(A) (X,P) +pSB1A3 (de-phosphorylated, E,P)
  • lac promoter/rbs(30)rep/oriV(6) + lac promoter/rbs(34)aada/oriT (A)
  • lac promoter/rbs(30)rep/oriV (6) (E,S) + lac promoter/rbs(34)aada/oriT(B) (X,P) +pSB1A3(de-phosphorylated, E,P)
  • oriV (E,S) + oriT (X,P) + pSB1A7 (E,P)
  • pSB1A7 self (did not de-phosphorylate) (E,P)


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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