Team:The University of Alberta/6 June 2008

(Difference between revisions)

Revision as of 21:06, 6 June 2008

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.
Jason designed the new ER1 biobrick

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!

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 *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.
+*Jason designed the new ER1 biobrick
 ==To Do==