Newcastle University Wetlab/3 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.
|
|
Wednesday 3rd September
- Today our aim was to transform our ligated plasmids into DH5-aplha E.coli competent cells.
- For each transformation, we added 100µl of DH5-alpha E.coli competent cells to 12.5µl of the ligation mixtures from yesterday.
- So in total we carried out 6 transformations, preparing them all on ice:
T1) Transforming pGFP-rrnB containing the 2.2kb fragment from pUC57-ncl08 into DH5-alpha
T2) Transforming pGFP-rrnB minus the 2.2kb fragment from pUC57-ncl08 into DH5-alpha
T3) Transforming pJWV021 containing the 2.2kb fragment from pUC57-ncl08 into DH5-alpha
T4) Transforming pJWV021 minus the 2.2kb fragment from pUC57-ncl08 into DH5-alpha
T5) Positive control: DH5-alpha cells plus pGFP-rrnB
T6) Negative control: DH5-alpha cells only
- The transformations were then plated onto antibiotic agar:
T1 : +spec
T2 : +spec (small and large volume plated)
T3 : +amp and +kan
T4 : +amp +kan
T5 : +spec
T6 : +spec