Team:NTU-Singapore/Notebook/12 July 2008



  

=Saturday 12 July= {|border="1" style="background-color:#ffffcc;" cellpadding="20"

Lab details

 * Choon Kit
 * Inoculation of colonies that were cloned on 11 July 2008
 * All agar plates that contained cells transformed with our new LacI-GFP showed positive growth. Each colony was distinct and selectable.
 * Necessary follow-ups were implemented. Inoculated 6 colonies off each plate, selected at random, into LBA broth. Each 5ml tubes of LBA + selected colony were incubated in a shaking incubator kept at 37oC for 16 hours.
 * Action to be followed up: Miniprep and EcoRI & PstI Digestion/ Agarose Gel Electrophoresis confirmation check on new LacI-GFP plasmids.
 * 1150bp band will be the correct band for LacI-GFP
 * 850bp band for GFP
 * 200bp band for LacI promoter
 * Transformation and Cloning of plasmids ligated on 11 July 2008
 * Plasmids that were ligated includes:
 * new part E7-AmpR vector (empty vector from GFP plasmid)
 * T7ptag-AmpR vector (empty vector from GFP plasmid)
 * A transformation and cloning into home-made Top10 competent cells were carried out. Instead of the usual practice to plate 250ul of LB/bacteria culture into one single LBA plate, it was divided into 2 plates, each containing approximately 125ul of bacteria culture. This modification was to encourage larger colony growth and enable selection at the follow up step easier. Cells that were given larger space to grow will develop into larger colonies.
 * Action to be followed up: Inoculation of E7-AmpR vector and T7ptag-AmpR vector.