Gels from yesterday:
Looks like:
Cofilin 1, 3, 4, 5 worked, need to do another digest.
LilrB2:TCSG4 1, 4, 5 might have worked. Strange extra band on top. Will do different digest.
LilrB2: YFPL none worked. Will miniprep more.
PirB:TCSG4 none worked.
PirB:YFP none worked.
To Do:
- Miniprep all the things
- Digest a couple of things
- Inoculate midi for pEXPR_hEF1a:LilrB2
Running gels:
HP1, LT1(B), LT1(X), LT4(B), LT4(X), LT5(B), LT5(X), YC1, YC3, YC4, YC5, LT4(B)
Cofilin worked! Sending all 4 for sequencing, 'cuz PCRs and mutations, ya know?
LilrB2:TCSG4 did not work. Will have to miniprep more.
Made glycerol cell stocks of all the cofilins, but due to some mistake, the labels got erased. So, unless they all worked, I'll just re-transform one that worked.
Making glycerol cell stocks of pEXPR_hEF1a: LilrB2 (L2 and L3)
Currently in stock: pENTR LilrB2, PirB (verified), pEXPR hEF1a: LilrB2/PirB (verified), pENTR_YFP:Cofilin (unverified)
Minipreped the new fusions (6-10)
Tried running gels for all of them. None worked.
But digested L2 and L3 again. They're fine! Will midi tomorrow.
8/11 comment: Drats! I should have posted a picture. What is "fine"? :'( I don't trust myself enough, given the confusion we are having now.