Team:NTU-Singapore/Notebook/16 July 2008
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(New page: =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 ef...) |
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*1240: MinElute PCR pufification for the above 7 mixtures (obtain 10ul each after purification). | *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. | *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. |
Revision as of 05:48, 24 July 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.