[Histonet] RNA isolation form stained slides

[Mark Tarango]

 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
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>