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Team:The University of Alberta/13 June 2008
From 2008.igem.org
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*Developed the Westerns after an overnight exposure. Blank yet again. James thought the problem was due to the development solution used - it was an old tube in the fridge that had been mixed months ago and was probably no good anymore. New solution was made and applied, film was exposed for 60 seconds and again, there were no results. This is odd because there were obvious thick bands of overexpressed proteins when we did the Coomassie staining. We will need to set up some cultures when we use the Ni-NTA columns so perhaps we will do them in duplicate and use one set for Westerns again. | *Developed the Westerns after an overnight exposure. Blank yet again. James thought the problem was due to the development solution used - it was an old tube in the fridge that had been mixed months ago and was probably no good anymore. New solution was made and applied, film was exposed for 60 seconds and again, there were no results. This is odd because there were obvious thick bands of overexpressed proteins when we did the Coomassie staining. We will need to set up some cultures when we use the Ni-NTA columns so perhaps we will do them in duplicate and use one set for Westerns again. | ||
*Digested I725021,I725022,I725025 and I725099 with xbal and pst | *Digested I725021,I725022,I725025 and I725099 with xbal and pst | ||
| - | *Did miniprep on Tom 2 night+ 1 day "overnight", did get a high yeild of DNA, so tried to run them on a gel. | + | *Did miniprep on Tom's 2 night+ 1 day "overnight", did get a high yeild of DNA, so tried to run them on a gel. |
==Lab Tip of the Day== | ==Lab Tip of the Day== | ||
When you do a digest, make sure you label the tubes clearly and be sure to include the name of the enzymes you used to do the digest. Since the BioBricks make use of multiple enzymes, it is important to know what has been digested with which, so the parts will fit together properly upon ligation. | When you do a digest, make sure you label the tubes clearly and be sure to include the name of the enzymes you used to do the digest. Since the BioBricks make use of multiple enzymes, it is important to know what has been digested with which, so the parts will fit together properly upon ligation. | ||
Revision as of 18:05, 13 June 2008
Today
- Developed the Westerns after an overnight exposure. Blank yet again. James thought the problem was due to the development solution used - it was an old tube in the fridge that had been mixed months ago and was probably no good anymore. New solution was made and applied, film was exposed for 60 seconds and again, there were no results. This is odd because there were obvious thick bands of overexpressed proteins when we did the Coomassie staining. We will need to set up some cultures when we use the Ni-NTA columns so perhaps we will do them in duplicate and use one set for Westerns again.
- Digested I725021,I725022,I725025 and I725099 with xbal and pst
- Did miniprep on Tom's 2 night+ 1 day "overnight", did get a high yeild of DNA, so tried to run them on a gel.
Lab Tip of the Day
When you do a digest, make sure you label the tubes clearly and be sure to include the name of the enzymes you used to do the digest. Since the BioBricks make use of multiple enzymes, it is important to know what has been digested with which, so the parts will fit together properly upon ligation.
