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Miniprepped more pENTR_LilrB2TCS
- Verify hEF1a: PirBTCS. Else pick more colonies. Picked more colonies
- Verify pENTR LilrB2TCS. Including LT16. Did not work
- Verify pEXPR_TREt:TEVpCofilin. Worked!
- Midiprep inoculate TREt:TEVpCofilin if it worked.
- Midiprep inoculate hEF1a: PirBTCS if it worked. Didn't work
- Sent pENTR_LilrB2 (04 2) for sequencing again...
Digest pEXPR_hEF1a: PirBTCS (1, 3, 4) with BsrGI, 2.1
Digest LT (21-25) with XbaI, XhoI, CS
Digest LT16 with NcoI, XhoI, CS
Digest pEXPR_TREt:TEVpCofilin with BsrGI, 2.1HL, LT 21, LT 22, LT 23, LT 24, LT 25, hPT1, hPT3, hPT4, LT16
HL, TTC 6, TTC 7, TTC 8, TTC 9, TTC 10 (image shows wrong names)
TREt:TEVpCofilin worked (6, 9, 10)! YES! Midiprep inoculate (9)!
LT's second band is still too low. Maybe the problem was with the PCR? Should we PCR again? After 25 colonies, I am not trying again.
hEF1a: PirBTCS did not work. Need to pick more