Team:NTU-Singapore/Notebook/22 July 2008



  

=Tuesday 22 July= {|border="1" style="background-color:#ffffcc;" cellpadding="20" Hung,Lu Chao, Min:

Ligation of E7 (vector) with Terminator (insert)

 * E7: cut with S/P for 2 hours
 * Terminator: cut with X/P for 2 hours
 * 1245: QIA PCR purification
 * 1300: gel loading (each sample into 3 lanes)
 * 1400: staining
 * 1430: Gel extraction, nanodrops. After that, we stored the digested DNAs (ready for ligation) at -20 fridge for ligation on Wednesday.

Ligation of lysis and LsrA into standard vector:

 * Cut Lysis,LsrA and GFP with E/P in Buffer 2, incubated for 2 hours.
 * 1345: QIA PCR purification
 * 1400: gel loading (each sample into 3 lanes)
 * 1500: staining
 * 1530: Gel extraction, nanodrops. After that, we stored the digested DNAs (ready for ligation) at -20 fridge for ligation on Wednesday.

Inoculation:

 * 3 lysis-terminator colonies transformed on Monday.
 * 3 E7-empty vector and 3 T7ptag-empt vector colonies.
 * 3 Terminator colonies (obtain more terminator plasmid for ligation).

Ligation of Lysis (I) and Terminator (V):

 * Do the same as Monday, in case all 3 colonies of Lysis-Term. are wrong.