User:Morgan L. Paull/Notebook/C.Dog V1.0 - T7 Promoter/2011/07/25: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 6: Line 6:
| colspan="2"|
| colspan="2"|
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==Picking colonies from Friday's full library transformation==
All look good, either red or green fluorescent under the light as appropriate, except for the following:
* G4 has some red colonies, it seems to be contaminated. It think this is ok however, because most colonies are green, and I can just pick a green one for the glycerol stocks. I initially picked a red one without thinking, and then picked a green one and put it in F4, ie same column but on its own row.
* R8.2 does not look red. I re-checked the sequencing and the sequencing looks fine, maybe it just is a very weak RBS, but it was not clear that it was working to me. I still picked it and put it in the normal place in the deep well culture plate for glycerol stocks
* G18.1 looked slightly red. However I have re-checked the sequencing results and they look fine.


I think that the way to test this moving forward to to inoculate G18.1 and R8.2 in LB + Kan + IPTG to see if they fluoresce properly under induced conditions.


Other than these two, the cultures all look great. The plate was not overgrown but had plenty of colonies in all cases. I can't find the plate of direct-from-transformation glycerol stocks/overnight cultures that Andy finished for me on Saturday, so I assume it is done and frozen, forming another backup for me.
 
==Making and checking the full library of constructs==
*inoculating (from last night's overnight cell culture) G18.1 and R8.2 in LB + Kan + IPTG to see if they fluoresce properly under induced conditions.
 


==Ligations from Friday==
==Ligations from Friday==
*Had several successful, visibly red R18 colonies! Horray! I picked them all, put them in EB and then put them in the fridge as well as starting overnight cell cultures. I will send them in for sequencing tomorrow.
*Sending in the picked colonies for sequencing, and making glycerol stocks (in individual tubes) from the overnight cell cultures made last night
 


<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Revision as of 11:23, 25 July 2011

C.Dog T7 Promoter Project <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>


Making and checking the full library of constructs

  • inoculating (from last night's overnight cell culture) G18.1 and R8.2 in LB + Kan + IPTG to see if they fluoresce properly under induced conditions.


Ligations from Friday

  • Sending in the picked colonies for sequencing, and making glycerol stocks (in individual tubes) from the overnight cell cultures made last night


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Morgan_L._Paull/Notebook/C.Dog_V1.0_-_T7_Promoter/2011/07/25&oldid=525675"