Newcastle University Wetlab/3 September 2008

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Newcastle University

GOLD MEDAL WINNER 2008

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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.

August
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September
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Wednesday 3rd September

Transformations

  • 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