Newcastle University Wetlab/3 September 2008
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(New page: {{:Team:Newcastle_University/WetLabCalendarHeader}} ==Wednesday 3rd September== Transformations * Today our aim was to transform our ligated plasmids into DH5-aplha E.coli competent...) |
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==Wednesday 3rd September== | ==Wednesday 3rd September== | ||
- | [[Transformations]] | + | [[Transforming into DH5α or TOP10 E. coli|Transformations]] |
* Today our aim was to transform our ligated plasmids into DH5-aplha E.coli competent cells. | * Today our aim was to transform our ligated plasmids into DH5-aplha E.coli competent cells. | ||
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'''T6)''' Negative control: DH5-alpha cells only | '''T6)''' Negative control: DH5-alpha cells only | ||
- | * The transformations were then plated onto antibiotic [[agar]]: | + | * The transformations were then plated onto antibiotic [[Making Agar Plates|agar]]: |
'''T1 :''' +spec | '''T1 :''' +spec |
Latest revision as of 16:35, 28 October 2008
Newcastle University
GOLD MEDAL WINNER 2008
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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.
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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