"
Newcastle University Wetlab/16 September 2008
From 2008.igem.org
Newcastle University
GOLD MEDAL WINNER 2008
| Home | Team | Original Aims | Software | Modelling | Proof of Concept Brick | Wet Lab | Conclusions |
|---|
Home >> Wet Lab >> Newcastle Wet Lab Journal
Wet lab work was carried out from 4 August to 19 September, Mondays to Fridays. Please click on a day to see the lab notebook.
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Tuesday 16th September
- Plasmid isolated from pGFPrrnB-ncl08 colonies 7 + 11 and pGFPrrnB-BB107 colonies 3 + 5 ON cultures.
- A Sample of each of the 4 isolated plasmids were restricted using 7.5μl plasmid and 0.5μl each enzyme in 15μl total volume:
- pGFPrrnB-ncl08 restricted with EcoRI + NheI
- pGFPrrnB-BB107 restricted with EcoRI + SpeI
Gel:
Expected band sizes:
* Lane 2: ~ 8.4kb and 2.2kb
* Lane 3: ~ 8.4kb and 2.2kb
* Lane 4: ~ 8.4kb and 3.2kb
* lane 5: ~ 8.4kb and 3.2kb
Lane 1: 1kb ladder
Lane 2: pGFPrrnB-ncl08 colony 7 restricted with EcoR1 and Nhe1
Lane 3: pGFPrrnB-ncl08 colony 11 restricted with EcoR1 and Nhe1
Lane 4: pGFPrrnB-BBa_I746107 colony 3 restricted with EcoR1 and Spe1
lane 5: pGFPrrnB-BBa_I746107 colony 5 restricted with EcoR1 and Spe1
As you can see from our gel, all 4 restrictions yielded the expected sized bands, indicating that the ligations had been successful. The remainder of the unrestricted plasmid from pGFPrrnB-ncl08 colony 7 was stored in the -20°C freezer ready to be sent off for sequencing. The two unrestricted pGFPrrnb-BBa_I746107 plasmid samples were then transformed into B.subtilis 168.
Agar plates made:
- pGFPrrnB-BBa_I746107 colony 3 (chloramphenicol)
- pGFPrrnB-BBa_I746107 colony 5 (chloramphenicol)
- B. subtilis 168 (negative control) (chloamphenicol)
