We check each cell mutations by a PCR and running on agarose gel , we obtained positive product with primers used for mutant colonies and some other mixture of cells, so seeded in a petri plates for more colonies and to confirm a mutation. The mutation is done by homologous recombination with a fragment of DNA containing a selectable marker URA 3. Then with specific primers for cells type wild and mutant cells I can see if there is mutation.
Ampicillin sterilize by filtering technique in a laminar flow and placing tubes in 20 to save and not to deteriorate the pregnant thawing and freezing all the time. The ampicillin comes in 1000x concentration (0.1 g / ml).
prepare a growth medium with:
1% yeast extract 2% peptone
2% Glucose Agar
2% (to prepare plates).
The image is of PCR and plasmid last week.
in the first well is the marker and through the well of the PCR first the positive and then negative (as there is nothing we can say that there was no contamination), and the last is that of the plasmid .
the person selling the marker delivery also with a paper showing how it will run and the bands that show sizes as base pairs. So we know how many base pairs have the PCR and plasmid .
in the first well is the marker and through the well of the PCR first the positive and then negative (as there is nothing we can say that there was no contamination), and the last is that of the plasmid .
the person selling the marker delivery also with a paper showing how it will run and the bands that show sizes as base pairs. So we know how many base pairs have the PCR and plasmid .
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