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Tet-inducible 3T3-L1 cells PowerPoint PPT Presentation


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LTR. GFP. CMV/TRE. PGK. Blast. LTR. Tet-inducible 3T3-L1 cells. RHKD lentivirus infection at <10%. Day 1. Seeding at 1000~2000 cells per well in 96-well plate, and induction with Dox. Day 2. Day 4. Induction of differentiation. Clone screening (total ~ 450,000). Day 10-12.

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Tet-inducible 3T3-L1 cells

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Tet inducible 3t3 l1 cells

LTR

GFP

CMV/TRE

PGK

Blast.

LTR

Tet-inducible 3T3-L1 cells

RHKD lentivirus infection at <10%

Day 1

Seeding at 1000~2000 cells per well in 96-well plate, and induction with Dox

Day 2

Day 4

Induction of differentiation

Clone screening (total ~ 450,000)

Day 10-12

Clone pickup and grow up with blasticidin, but without Dox

Re-differentiation with and without Dox

Gene identification

shRNA verification


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