[Histonet] RNA isolation form stained slides

["E. Wayne Johnson 朱稳森"]

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"<
>> marktarango <@t> gmail.com<javascript:_e({}, 'cvml',
>> 'marktarango <@t> 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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>>>        
>>      
>