IGEM:UNAM LCG/2009/Notebook/Hydrobium etli/2011/09/23
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HydA and Hyd G digestionAbstractI made a double digestion from HydA and HydG with EcoRI and PstI to extract them from the plasmid in which they were sent to us by Mr. Gene. The objective is to ligate this parts with the plasmid pSB1C3 to send them to the registry. Apparently something went grong in the digestion of HydA. DetailsI prepared 4 sample for each of the parts (hydA and hydG) so I made 4 digestions for each part but this digestions were replicas, all had the same ammounts of components. Digestion details
ResultsI ran a gel to see if the digestions were well.
Gel imageLanes: 1. HydA 1 2. HydA 2 3. HydA 3 4. HydA 4 5. HydG 1 6. HydG 2 7. HydG 3 8. HydG 4 9. Ladder fermentas 10. pSB1C3 11-rest. empty Gel analysisThe double digestion of pSB1C3 was made by Gustavo, He put it in this gel to see the concentration and calculate the ammaunt he will need to do a ligation, I will use this same sample to do the ligation in the next step. It apperas that HydA digestion did not work out because there is a single band, I have to ask Pae if this is the expected size anyway. Besides Helena is tring to take off the tag. I am not sure if the diggestions for HydG were well because I do not know what are the expected sizes, Pae will analize it and tell me. |