McGill/September
From 2007.igem.org
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== September 2007 == | == September 2007 == | ||
| - | ===September | + | ===September 4=== |
| + | Performed a screening digest of E0434 & I5611 after a midiprep. | ||
| + | Results: Not so good, however, linear cuts showed that the DNA was present, so ligation will go ahead. | ||
| + | |||
| + | ===September 5=== | ||
| + | *Transformed I5611 & E0434 again in Top 10 cells. Used a 2 min heat shock time, and plated on Amp/Kan (E0434) and Amp (I5611) plates. | ||
| + | *Did a large scale digest I5611 & E0434. However, there might not have been enough BsrGI enzyme added, hence cuts were not clear and pronounced. Digest must be performed again. | ||
| + | |||
| + | ===September 7=== | ||
| + | *Ligated linear I5611 & E0434. Left overnight at room temperature instead of at 37C in a water bath. | ||
| + | *Then performed a screening digest of I5611 & E0434. This proved unsuccessful again. | ||
| + | |||
| + | ===September 11=== | ||
| + | *Got excellent colonies for all ligation plates (used- Insert:Vector [1:1, 1:2, 1:3, 1:4, 1:5]x2). Each plate had ~25 colonies. | ||
| + | *Seeded 12 colonies from each plate, in 1X LB with Amp/Kan resistance. Thus, seeded 60 colonies in total, the culture tubes were then left in the shaker/incubator at 5.30pm, to be kept overnight. | ||
Revision as of 04:14, 25 October 2007
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Contents |
September 2007
September 4
Performed a screening digest of E0434 & I5611 after a midiprep. Results: Not so good, however, linear cuts showed that the DNA was present, so ligation will go ahead.
September 5
- Transformed I5611 & E0434 again in Top 10 cells. Used a 2 min heat shock time, and plated on Amp/Kan (E0434) and Amp (I5611) plates.
- Did a large scale digest I5611 & E0434. However, there might not have been enough BsrGI enzyme added, hence cuts were not clear and pronounced. Digest must be performed again.
September 7
- Ligated linear I5611 & E0434. Left overnight at room temperature instead of at 37C in a water bath.
- Then performed a screening digest of I5611 & E0434. This proved unsuccessful again.
September 11
- Got excellent colonies for all ligation plates (used- Insert:Vector [1:1, 1:2, 1:3, 1:4, 1:5]x2). Each plate had ~25 colonies.
- Seeded 12 colonies from each plate, in 1X LB with Amp/Kan resistance. Thus, seeded 60 colonies in total, the culture tubes were then left in the shaker/incubator at 5.30pm, to be kept overnight.
