Team:NTU-Singapore/Notebook/22 July 2008
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(Difference between revisions)
m (New page: =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) |
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*E7: cut with S/P for 2 hours | *E7: cut with S/P for 2 hours | ||
*Terminator: cut with X/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. |
Revision as of 07:31, 24 July 2008
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.