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17/09/13
From 2013.igem.org
(Difference between revisions)
(Created page with "==Results from previous day== *Background control plate had growth, which was not expected. This suggests that pSB1C3 backbone religated. *It was also thought that the Tod genes...") |
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*For digestion of backbone, already digested backbone from 01/08/2013 was used, which was digested with EcoRI and PstI. | *For digestion of backbone, already digested backbone from 01/08/2013 was used, which was digested with EcoRI and PstI. | ||
| - | == | + | ==Incubation of bacteria from the plates of previous day for mini prep== |
| + | *12 single colonies were picked from each plate and put into 15ml of broth | ||
| + | *36 tubes were left incubating overnight at 37C | ||
| + | |||
| + | ==Sending pGEM-T vector with Tod insert clones to sequencing by PNACL== | ||
| + | |||
| + | ==Digestion of new fusion PCR Tod genes== | ||
| + | *Digestion with EcorI-HF and PstI-HF using Cutsmart buffer | ||
| + | {|border=1 | ||
| + | |Sample||DNA||Buffer||Water||EcorI-HF||PstI-HF | ||
| + | |- | ||
| + | |Tod X||10.1ul||3ul||15.9||0.5||0.5 | ||
| + | |- | ||
| + | |Tod F||13.4ul||3ul||12.6||0.5||0.5 | ||
| + | |- | ||
| + | |Tob B||14.7ul||3ul||11.3||0.5||0.5 | ||
| + | |} | ||
Revision as of 10:16, 18 September 2013
Contents |
Results from previous day
- Background control plate had growth, which was not expected. This suggests that pSB1C3 backbone religated.
- It was also thought that the Tod genes used were not from the PCR fusion experiment.
- Same experiment done again, with a new set of Tod genes from a PCR fusion.
- For digestion of backbone, already digested backbone from 01/08/2013 was used, which was digested with EcoRI and PstI.
Incubation of bacteria from the plates of previous day for mini prep
- 12 single colonies were picked from each plate and put into 15ml of broth
- 36 tubes were left incubating overnight at 37C
Sending pGEM-T vector with Tod insert clones to sequencing by PNACL
Digestion of new fusion PCR Tod genes
- Digestion with EcorI-HF and PstI-HF using Cutsmart buffer
| Sample | DNA | Buffer | Water | EcorI-HF | PstI-HF |
| Tod X | 10.1ul | 3ul | 15.9 | 0.5 | 0.5 |
| Tod F | 13.4ul | 3ul | 12.6 | 0.5 | 0.5 |
| Tob B | 14.7ul | 3ul | 11.3 | 0.5 | 0.5 |
