请教stromal vascular fraction (SVF) from white adipose tissue 的几个问题# Biology - 生物学
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最近要提取从白色脂肪里面提取SVF,根据protocol我确实看到了pellet,用TRI-
reagent也提取到了mRNA,nanodrop测出的浓度有800ng/ul,但是我试了cyclophilin,
gapdh,18s几个内参,都测不到CT值。已经重复几次了,郁闷的不行。请教各位,有什
么建议么?
SVF protocol
1)isolate WAT,
2) Incubate tissue @ 37C for 45min with shaking @ 80 rpm (Collagenase I,
1mg/ml)
3)300g for 10min
4) Dissolve pellet in Tri-reagent.
reagent也提取到了mRNA,nanodrop测出的浓度有800ng/ul,但是我试了cyclophilin,
gapdh,18s几个内参,都测不到CT值。已经重复几次了,郁闷的不行。请教各位,有什
么建议么?
SVF protocol
1)isolate WAT,
2) Incubate tissue @ 37C for 45min with shaking @ 80 rpm (Collagenase I,
1mg/ml)
3)300g for 10min
4) Dissolve pellet in Tri-reagent.