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.
TaqMan PreAmp plates
| Plate
|
Genes
|
| #1
|
Bv8, Id3, EGF, Tie1
|
| #2
|
MMP12, CXCR4, MMP9, CX3CR1
|
| #3
|
YM1, CD115, Arg1, Tie2
|
| #4
|
GAPDH, Angpt1, Angpt2, MMP2
|
| #5
|
SDF, CD62L, VEGFR2, B2m
|
| #6
|
VEGFR1, bFGF, RelB, MafB
|
| #7
|
|
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:
- 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
EB1 (Tris-HCl) 70 ul, 1' RT, 1' 13k rpm
|
TEMs gene expression profiling
Main project page
