Team:NTU-Singapore/Notebook/16 July 2008



  

=Wednesday 16 July= {|border="1" style="background-color:#ffffcc;" cellpadding="20" DNA: 1000ng (around 5ul) Enzyme1: 1 ul Enzyme2: 1 ul Buffer: 5 ul BSA: 0.5 ul H2O: 37.5 ul Total: 50
 * 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:
 * E7 and T7ptag with E/P in buffer 2 for 1.5 hours. For all the digestions, the amounts used are:
 * 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.