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Team:University of Ottawa/31 July 2008
From 2008.igem.org
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:<li> Another ligation of PTP2 insert with pSSA42 vector was performed with 3:1 ratio, 6:1 ratio, Vector with ligase, vector without ligase and H2O control. | :<li> Another ligation of PTP2 insert with pSSA42 vector was performed with 3:1 ratio, 6:1 ratio, Vector with ligase, vector without ligase and H2O control. | ||
:<li> 2 hr ligation was unsucessful, overnight ligation was usccessful for both 3:1 and 6:1. | :<li> 2 hr ligation was unsucessful, overnight ligation was usccessful for both 3:1 and 6:1. | ||
| + | '''Transformation''' | ||
| + | :<li> Transformed ligation product into competent cells - 3 different plates -3:1, 6:1 and control. | ||
Revision as of 19:10, 7 August 2008
Today in the Lab
Chris
Microwave Digestion Experiment
- Following protocol outlined online, tested a microwave protocol to hasten digestion period.
- Digested DQ232597 with HindIII. Ran five samples: 5 seconds in micro, 10 seconds, 15 seconds, 10 seconds in and 2 minutes out (four times), water control, water control 10 seconds in microwave and 2 minutes out, 1 hr traditional digest.
- Denatured HindIII at 80 C for 20 minutes, then ran products on a gel.
- Results promising; however, more experimentation needed to confirm validity of this protocol
Matt
Ligation
- Another ligation of PTP2 insert with pSSA42 vector was performed with 3:1 ratio, 6:1 ratio, Vector with ligase, vector without ligase and H2O control.
- 2 hr ligation was unsucessful, overnight ligation was usccessful for both 3:1 and 6:1.
Transformation
- Transformed ligation product into competent cells - 3 different plates -3:1, 6:1 and control.
