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Re: 检测基因表达变化时,RT-PCR和Northern blot首选哪个?
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Re: 检测基因表达变化时,RT-PCR和Northern blot首选哪个?# Biology - 生物学
s*e
1
clonist //hehe, take it easy.
generally, RT-pcr is the most sensitive assay to RNA, although sometimes
it cause pseudo positive.
However, RT-PCR can't detect isoforms, or alternative splicing products.
And because of its pseudo positive.
a northern is necessary.
However, S1 mapping of nuclease protection assay is roughly 20 fold more
sensitive than northern. Although these two methods also can't give
you the information of the full length or some isoforms' information.
all in all, these assays a
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M*e
2
if you are look at splicing events and you definitely would
like to use northern blot, though technically it might be
difficult. RTPCR is more sensitive for quantitation of total
messages, or you can actually design primers for alternative
splicing products to specifically measure them in a total
population of mRNAs. i like RTPCR since it is quick to roughly
estimate changes for steady level of RNA. and sometimes i do
semiquantitative RTPCR, which means that i stop the RXN before
it reaches the
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