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EPF-Lausanne/18 August 2008
From 2008.igem.org
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=Molecular Biology= | =Molecular Biology= | ||
| - | == | + | ==Checking all the parts== |
| - | We made a gel to make sure that the PCR of some Biobricks, especially the small ones, worked. As they worked well, we can directly use them for the digestion, they will be in excess and we can therefore be sure that the insertion in the other Biobrick's plasmid will work relatively well. | + | We made a gel to make sure that the PCR of some Biobricks (PCR'd on friday), especially the small ones, worked. As they worked well, we can directly use them for the digestion, they will be in excess and we can therefore be sure that the insertion in the other Biobrick's plasmid will work relatively well. |
| + | |||
| + | Result: | ||
| + | All fragments are visible at the right place, although small fragments almost run out of the gel (100V, 45') and are just barely visible. Maybe PCR didn't work very well for those. | ||
| + | |||
| + | ==Ligations== | ||
| + | |||
| + | * We have the inserts R0071 and I1466 from friday, we redigest (adding Antarctic Phosphatase after this) and purify by gel the two corresponding vectors (ie B0034 and I14033. After this we do the ligation and transformation, letting the bacteria grow overnight. | ||
| + | |||
| + | * We start also with the third ligation R0040 / B0034 (vector). We digest them and run the gel. | ||
Revision as of 07:34, 19 August 2008
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Molecular Biology
Checking all the parts
We made a gel to make sure that the PCR of some Biobricks (PCR'd on friday), especially the small ones, worked. As they worked well, we can directly use them for the digestion, they will be in excess and we can therefore be sure that the insertion in the other Biobrick's plasmid will work relatively well.
Result: All fragments are visible at the right place, although small fragments almost run out of the gel (100V, 45') and are just barely visible. Maybe PCR didn't work very well for those.
Ligations
- We have the inserts R0071 and I1466 from friday, we redigest (adding Antarctic Phosphatase after this) and purify by gel the two corresponding vectors (ie B0034 and I14033. After this we do the ligation and transformation, letting the bacteria grow overnight.
- We start also with the third ligation R0040 / B0034 (vector). We digest them and run the gel.
