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Electroporate pBR322+GFP(LVA) into
Second Attempt: Quantitate the Constitutive Expression Level of GFP fused to pBR322 P1 Promoter
Create pBR322+GFP(LVA)
Digest pBR322 and pGFP(LVA) with 2 Enzymes Each
Gel Purify Digested pBR322 and GFP(LVA)
Anneal XbaI-EcoRI Adapter
Ligate pBR322, GFP(LVA), adapters
Gel Purify Ligation of pBR322 + GFP(LVA)
Quantitate expression levels of GFP on individual cells using FACS
Obtain calibration curve with fluorescent beads
Stain cells to enable distinguishing cells from debris
Run stained cells through cytometer and gather data
Analyze data
Ron Weiss
Wed Feb 10 15:48:08 EST 1999
