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16/09/13
From 2013.igem.org
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==Running the backbone and RFP on gel== | ==Running the backbone and RFP on gel== | ||
*on 1% gel to check if the gel extraction worked. | *on 1% gel to check if the gel extraction worked. | ||
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| + | ==Digestion of the pGEM plasmid extracted on the [[13/09/13]]== | ||
Revision as of 12:30, 16 September 2013
Contents |
Digesting pSB1C3 backbone
- Digesting for ligation with the already digested Tod operon PCR products
- Following the iGEM official protocol
- Using EcoRI-HF and PstI-HF
- Exluding the DpnI from the master mix and adjusting the volume of water accordingly
- Protocol can be found at: [1]
Digestion of the TOD genes PCR products
The PCR products of TODX, TODF and TOBG were digested with the enzymes EcoRI-HF and PstI-HF.
Protocol
| Sample | DNA volume (ul) | cutsmart buffer (ul) | EcoRI (ul) | PstI (ul) | dH2O (ul) |
| TODX | 8.7 | 2 | 0.5 | 0.5 | 8.3 |
| TODF | 8.3 | 2 | 0.5 | 0.5 | 8.7 |
| TOBG | 8 | 2 | 0.5 | 0.5 | 9 |
- The samples were incubated for 1 hour at 37C.
- Heat kill the enzymes at 80C for 20 minutes.
Running the backbone and RFP on gel
- on 1% gel to check if the gel extraction worked.
