User:Ferdinando Pucci/Notebook/TEMs profile/2008/04/28

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] TEMs gene expression profiling
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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TaqMan PreAmp

 * cDNA from 4 sortings, Mono6-9, CD115+Gr1+ inflammatory monocytes and CD115+Gr1- resident monocytes, 12.5ul each
 * 28 assay pooled: Angpt1, Angpt2, B2m, CD163, SDF1, EDG1, bFGF, VEGFR1, IGF1, VEGFR2, MafB, RelB, MMP2,9,12, Tie1, Tie2, LYVE1, EGF, Id3, Bv8, CXCR4, CX3CR1, CD62L, Arg1, YM1, CD115, GAPDH. 1:100
 * cDNA done with Oligo-dT and poliA as carrier

N202 injection
5*106 cells/mouse, 50 mice and 4 PGK-GFP. Kept 3 15-cm plates in line.

IFN exp3
RNA extraction from organs (Bra, Lun, Hea, Liv, Gut, Spl, Kid, BM) of mice BMT without tumors, long term. DNA+RNA from BM: EB1 (Tris-HCl) 70 ul, 1' RT, 1' 13k rpm
 * RNeasy mini plus kit
 * Lysate without EtOH on column, 30'' max rpm
 * EtOH 70%, 1:1, RNA as the others (with DNase treatment)
 * AW1 500ul
 * AW2 500ul 2' 11k rpm, change tube
 * 1' 13k rpm


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