Todo Today:
- Miniprep LRs
- Digest LRs
- Also digest pEXPR_hEF1a: PirBYFP
- Transform PirB-YFP LRs (maybe do treatment's one too...). Nope.
- PCR LilrB2 for YFP and TCSG4
- Run: Digests, PCRs
- Cut out PCR bands if worked.
- Midi inoculate if LilrB2 LRs worked.
- LR pENTR_PirB:YFP to hEF1a and TREt
PCRing at 53 degress, 40 seconds.
Digesting with:
pEXPR_hEF1a: LilrB2 XhoI, CS
pEXPR_TREt: LilrB2 XbaI, CS
pEXPR_hEF1a: PirB_YFP ApaI, CS
Order:
HL, pEXPR_hEF1a:LilrB2 (1-4), pEXPR_hEF1a:LilrB2 C, p(??)_PirB_YFP, PCR_LilrB2(Y) test, PCR_LilrB2(T) test
HL, pEXPR_TREt:LilrB2 (1-4), pEXPR_TREt:LilrB2 C, PCR_LilrB2(Y), PCR_LilrB2(T)
Both LilrB2 pEXPRs worked!
The PCR has a band in the right length area. Will gel extract.'
PirB looks like it's the pENTR not the pEXPR.