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Team:The University of Alberta/6 June 2008
From 2008.igem.org
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==Today== | ==Today== | ||
*We got the new reverse primers for Purple Russian and Tryp in today. We set up PCR using these reverse primers with the forward primers for Purple Russian and Tryp that we got last week (using the PFU polymerase protocol). | *We got the new reverse primers for Purple Russian and Tryp in today. We set up PCR using these reverse primers with the forward primers for Purple Russian and Tryp that we got last week (using the PFU polymerase protocol). | ||
| + | *These were then digested with XbaI and PstI. We also digested J61003 so we can ligate them together, transform them and check for expression. If it works, we can then cut them out using XbaI and PstI again, and put them into I0500 so we can get rid of the promotor between the EcoRI and XbaI sites. | ||
*Some of the O/Ns we set up yesterday didn't grow; those that did were 25, 35 and thiolase. We have set up overday cultures of the ones that did not grow. | *Some of the O/Ns we set up yesterday didn't grow; those that did were 25, 35 and thiolase. We have set up overday cultures of the ones that did not grow. | ||
*Tom has made another two polyacrylamide gels; we will run the crude and soluable proteins from 25, 35 and thiolase on these. | *Tom has made another two polyacrylamide gels; we will run the crude and soluable proteins from 25, 35 and thiolase on these. | ||
Revision as of 19:56, 6 June 2008
Today
- We got the new reverse primers for Purple Russian and Tryp in today. We set up PCR using these reverse primers with the forward primers for Purple Russian and Tryp that we got last week (using the PFU polymerase protocol).
- These were then digested with XbaI and PstI. We also digested J61003 so we can ligate them together, transform them and check for expression. If it works, we can then cut them out using XbaI and PstI again, and put them into I0500 so we can get rid of the promotor between the EcoRI and XbaI sites.
- Some of the O/Ns we set up yesterday didn't grow; those that did were 25, 35 and thiolase. We have set up overday cultures of the ones that did not grow.
- Tom has made another two polyacrylamide gels; we will run the crude and soluable proteins from 25, 35 and thiolase on these.
- Jason digested Blue Ox with PstI and XbaI; this will allow us to put it into I0500 so we get rid of the 200bp promotor and have a 100% totaly proper BioBrick.
To Do
- Check to see if the Ni columns have arrived so we can purify the His-tagged proteins from the butanol stuff
- Do westerns on the crude and soluable PAGE gels
- Check out how our NOT Gate is going to work
- Build biobricks!
