Team:Hawaii/Construction of pRL1383a
From 2008.igem.org
Projects | Events | Resources | ||
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Sponsors | Experiments | Milestones | Protocols | |
Notebook (t) | Meetings (t) |
Contents |
Ligation of Parts
- The BioBrick parts of pRL1383a are to be ligated in a series of experiments.
Methods
- Round 1 of Restriction Digest and Ligation
Name | Digest 1 | Digest 2 | Ligation to | Product |
---|---|---|---|---|
mob | XbaI & PstI | n/a | BBa_B0024 | TT-mob |
rep | XbaI & PstI | n/a | BBa_R0010 | Plac-rep |
oriV | XbaI & PstI | n/a | pSB1A3 | pSB1A3::oriV |
aadA region | XbaI & PstI | n/a | BBa_R0010 | Plac-aadA |
oriT | XbaI & PstI | n/a | pSB1A3 | pSB1A3::oriT |
pSB1A3 | XbaI & PstI | n/a | see oriT and oriV | see oriT and oriV |
BBa_B0024 | SpeI | PstI | see mob region above | see mob region above |
BBa_R0010 | SpeI | PstI | see rep and aadA region above | see rep and aadA region above |
- In gel purification of ligation products
- Round 2 of Restriction Digest and Ligation
Name | Digest 1 | Digest 2 | Ligation to | Product |
---|---|---|---|---|
TT-mob | EcoRI | SpeI | BBa_B0015 | TT-mob-TT |
Plac-aadA | EcoRI | SpeI | BBa_B0015 | Plac-aadA -TT |
BBa_B0015 | EcoRI | XbaI | see mob and aadA above | see mob and aadA above |
- Transformation
- Verification with Colony PCR
Results
Discussion
- What was learned and how to do future experiments differently.