Berk2006-Sequences

To Do

Ogel MC600u x M1/P1 (2 clones), pBACr-90* x MG984, pMF19, MG1655 x MC600u/P1 (2 clones plus dust?)

 

Comp MGu x UG874

 

sub pSB3C6-Bca1020 (for r0040)

Sub pMP908

 

Sub new locks

Sub new promoter J23119

 

Grow to assay J23066, x locks (5:30)

Stock, seq 62, M1, UG874

Outgrow UG874 x MC600u

 

Riboregulators

To do: keys that anneal further upstream, repairing the rbs in lock, cytometry on the above.

Make pSB3C6-I13521

 

Conjugation

Screen the trbC mutants by PCR/phenotype, clear pKD46

oriTf mutant x 1064, test phenotype

Rlam traG mutant, screen colonies

trbC/traG complementation plasmids (Pcon-rbs is good, maybe need to reprep)

 

 

 

 

 

LB + 5 mM CaCl2 and 0.2% glucose

LB + 100 mM MgSO4 + 5 mM CaCl2

200 µL 1 M Na-Citrate (pH 5.5)

chloroform

 

Berk prop/email adam

Meet w/ Wendell,Shaffer,other bioE

 

Solid phase on plate

Sequence pCP20, clone it into new pMP plasmid

UCSF Safety

K99

Mouse serum/Balb/c

Venter

 

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Inject BO828/pBACr-CyoWbb / pAC-TetInv / pBR-GFP into tumors, tetR library, fdhF-tetR inverter

 

Email Jamie for protocols

Email Jack about tailvein injections