Team:The University of Alberta/6 June 2008

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Revision as of 22:46, 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.

EDIT:Digest went for ~2 hours; the Tryp and Purple Russian parts were ligated into J61003 and the ligation was left at room temperature overnight. Chris will come in ~noon on Saturday to move them into the freezer (unless there is a volunteer coming in on Saturday, in which case, contact him (434-0508/cwk AT ualberta DOT ca)

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
Did Westerns on the crude and soluable proteins of the butanol parts. Put them in the fridge until Monday; we will do the antibody detection then.
Stained the PAGE gel that Tom ran; will let it destain in the fridge over the weekend.

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!

@@ Line 1: / Line 1: @@
 ==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.
+*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.<br>
+'''EDIT:'''Digest went for ~2 hours; the Tryp and Purple Russian parts were ligated into J61003 and the ligation was left at room temperature overnight. Chris will come in ~noon on Saturday to move them into the freezer (unless there is a volunteer coming in on Saturday, in which case, contact him (434-0508/cwk AT ualberta DOT ca)
 *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
+*Did Westerns on the crude and soluable proteins of the butanol parts. Put them in the fridge until Monday; we will do the antibody detection then.
+*Stained the PAGE gel that Tom ran; will let it destain in the fridge over the weekend.
 ==To Do==