z***x
发帖数: 80 | 1 I have no experience of isolating RNA from cartilage but need to do it soon.
Cartilage contains a lot of proteoglycans and glycosaminoglycans (GAGs). I
first homogenized cartilage into powders by freeze-milling. I then tried
guanidine and phenol to dissolve the cartilage powder but it was not quite
efficient. Most of the 100mg of cartilage powder was not dissolved in 2 ml
guanidine/phenol (Qiagen tissue lysis reagent).
The other question is that RNA and GAGs are both negatively charged polymers
. I think they will co-purify on the RNA binding column. I suspect they will
co-precipitate with ethanol/IPA precipitation although I haven't tried to
do it. If the purified RNA sample contains GAGs, will I get false reading
for OD 260/280? Will the GAG contamination interfere microarray analysis?
I need about 2ug of total RNA from cartilage,
so how much cartilage should I start with?
which method to dissolve the tissue?
Will GAGs interfere with OD260/280 reading?
Will GAGs interfere with microarray analysis?
Thanks. | d******e
发帖数: 351 | 2 still worried about this? just emailed you. hehe, 帮顶一个 |
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