Team:NTU-Singapore/Notebook/22 July 2008
From 2008.igem.org
Contents |
Tuesday 22 July
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.