I read one paper or abstract recently where sections were made at 50
microns then used
successfully for RT-PCR. I am pretty sure that it was PRRS virus that
they were pursuing.
On 3:59, Mark Tarango wrote:
I would try getting more sections from LCM and extracting into 7.5 uL, if
that volume will be enough to for your PCR reactions.
On Friday, November 2, 2012, Vanessa Orsini wrote:
With the kit I'm extracting in 10ul....the problem is that I have too use
few stained cells isolated with the LCM...so even if I increase the number
of slides I'll never have a lot of material...
Inviato da iPhone
Il giorno 2 nov. 2012, alle ore 16:44, "Mark Tarango"<
marktara...@gmail.com<javascript:_e({}, 'cvml',
'marktara...@gmail.com');>> ha scritto:
Have you tried using more sections during extraction? Can you extract
into a smaller volume?
On Friday, November 2, 2012, Vanessa Orsini wrote:
Hello,
I need to extract RNA for a RT-PCR after Laser Micro
Dissection on xGal stained slides.
I tried using sections from unfixed frozen organs. I fixed the
sections in EtOH70% for 10min and then I stained them with xGal for 3h at
37°C.
After air drying I cut out with the LCM and extract RNA with the PicoPure
kit
from Applied Biosystem. So far I didn’t manage to get enough RNA.
I tried to add RNase inhibitors to all the solutions but it
didn’t help.
Any idea/suggestion?
Do someone think it would be better to do a LacZ antibody staining
on FFPE sections and extract RNA with an appropriate kit? The RNase would
they
be less active?
Thank in advance for any help you can give me J Vanessa
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