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.