User:Morgan L. Paull/Notebook/C.Dog V1.0 - T7 Promoter/2011/07/24

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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.

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.