Team:NTU-Singapore/Notebook/16 July 2008

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=Wednesday 16 July=
=Wednesday 16 July=
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*Morning: We carried out several digestion:
*Morning: We carried out several digestion:
**GFP with E/P in buffer 2 for 1 hour; in buffer 2 for 2 hours; and in EcoRI buffer for 2 hours (this aims to compare the efficiency of buffer 2 with EcoRIbuffer for E/P double digestion; as well as the optimal digestion time).
**GFP with E/P in buffer 2 for 1 hour; in buffer 2 for 2 hours; and in EcoRI buffer for 2 hours (this aims to compare the efficiency of buffer 2 with EcoRIbuffer for E/P double digestion; as well as the optimal digestion time).
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*1650: gel extraction. After this, the digested DNAs were put in -20 fridge for ligation on Thursday.
*1650: gel extraction. After this, the digested DNAs were put in -20 fridge for ligation on Thursday.
*1520: digestion for gel check of 12 samples of Fe-GFP inoculated yesterday. However,no colony was found correct.
*1520: digestion for gel check of 12 samples of Fe-GFP inoculated yesterday. However,no colony was found correct.
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Revision as of 12:14, 25 October 2008

Wednesday 16 July

  • Morning: We carried out several digestion:
    • GFP with E/P in buffer 2 for 1 hour; in buffer 2 for 2 hours; and in EcoRI buffer for 2 hours (this aims to compare the efficiency of buffer 2 with EcoRIbuffer for E/P double digestion; as well as the optimal digestion time).
    • GFP with X/P in buffer 3 for 2 hours.
    • E7 with X/P in buffer 3 for 2 hours.
    • E7 and T7ptag with E/P in buffer 2 for 1.5 hours.
      For all the digestions, the amounts used are: 
DNA: 1000ng (around 5ul)
Enzyme1: 1 ul
Enzyme2: 1 ul
Buffer: 5 ul
BSA: 0.5 ul
H2O: 37.5 ul
Total: 50
  • 1240: MinElute PCR pufification for the above 7 mixtures (obtain 10ul each after purification).
  • 1650: gel extraction. After this, the digested DNAs were put in -20 fridge for ligation on Thursday.
  • 1520: digestion for gel check of 12 samples of Fe-GFP inoculated yesterday. However,no colony was found correct.
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