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Team:University of Ottawa/21 July 2008
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Djedrysiak (Talk | contribs) (New page: __TOC__ ==Today in the lab== '''Dan''' :'''Gel of sample 2T''' ::<li>For some odd reason all of the DNA stayed in the well... Could be some sort of salt contamination from NEB buffer 1, or...) |
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:'''Gel of sample 2T''' | :'''Gel of sample 2T''' | ||
::<li>For some odd reason all of the DNA stayed in the well... Could be some sort of salt contamination from NEB buffer 1, or a very large autoligation product. I will test the former by doing PCR cleanup and switching to ligation buffer and then run the gel again. | ::<li>For some odd reason all of the DNA stayed in the well... Could be some sort of salt contamination from NEB buffer 1, or a very large autoligation product. I will test the former by doing PCR cleanup and switching to ligation buffer and then run the gel again. | ||
| - | '''Digestion of 1A 0B and both S and D''' | + | :'''Digestion of 1A 0B and both S and D''' |
::<li> The above DNA fragments were digested with PstI and SphI in Neb buffer 1, enzymes were denatured after an hour | ::<li> The above DNA fragments were digested with PstI and SphI in Neb buffer 1, enzymes were denatured after an hour | ||
::<li> PCR cleanup was performed on the digested products, eluted with 50 uL water. | ::<li> PCR cleanup was performed on the digested products, eluted with 50 uL water. | ||
| - | '''Ligation''' | + | :'''Ligation''' |
::<li> Ligation of the DNA obtained from PCR cleanup was run overnight at 16 C | ::<li> Ligation of the DNA obtained from PCR cleanup was run overnight at 16 C | ||
Revision as of 14:34, 22 July 2008
Contents |
Today in the lab
Dan
- Gel of sample 2T
- For some odd reason all of the DNA stayed in the well... Could be some sort of salt contamination from NEB buffer 1, or a very large autoligation product. I will test the former by doing PCR cleanup and switching to ligation buffer and then run the gel again.
- Digestion of 1A 0B and both S and D
- The above DNA fragments were digested with PstI and SphI in Neb buffer 1, enzymes were denatured after an hour
- PCR cleanup was performed on the digested products, eluted with 50 uL water.
- Ligation
- Ligation of the DNA obtained from PCR cleanup was run overnight at 16 C
