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Team:Hawaii/Notebook/2008-10-15
From 2008.igem.org
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(margaret) |
(→Construction of secretion device (cont.)) |
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::* Used 1 μl of J33207/pRL1383a 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 | :::* Plated 25 μl, 50 μl, 75 μl, and 100 μl plates in hopes of isolating single colonies | ||
| + | |||
| + | :<strong> Krystle </strong> | ||
| + | :* 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=== | === Ligation=== | ||
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
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
