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FACS Sample Prep from whole brain tissue (mouse)


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#1 noodle

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Posted 16 March 2009 - 09:48 AM

I'm looking for a protocol for sample preparation for FACS from whole brain. I'm using mouse pups at postnatal day 1, and would like to be able to extract RNA for gene arrays after running FACS.

If anyone has any advice, it'd be greatly appreciated!

Thanks! :unsure:

#2 CKtong

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Posted 16 March 2009 - 06:21 PM

I'm looking for a protocol for sample preparation for FACS from whole brain. I'm using mouse pups at postnatal day 1, and would like to be able to extract RNA for gene arrays after running FACS.

If anyone has any advice, it'd be greatly appreciated!

Thanks! :)

i suggest you split the cell for your FCM and RNA, it shud be enuf. After flow you my have the issue that you RNA extraction may no have enuf sample or contamination......
There is not always you can kill two bird by one stone.

#3 WOW

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Posted 19 March 2009 - 10:17 PM

I'm looking for a protocol for sample preparation for FACS from whole brain. I'm using mouse pups at postnatal day 1, and would like to be able to extract RNA for gene arrays after running FACS.

If anyone has any advice, it'd be greatly appreciated!

Thanks! ^_^

i suggest you split the cell for your FCM and RNA, it shud be enuf. After flow you my have the issue that you RNA extraction may no have enuf sample or contamination......
There is not always you can kill two bird by one stone.


If you want to extract RNA from a subpopulation of cells, which only could be obtained from FACS, you would have no choice. If you simply analyze the cell mass for a control of cell status then extract RNA for array, then advice from CKtong would be nice.
FACS would cause some damage to your cell, which depends on the expertise of your FACS facility. If you gonna sort neurons, you should well plan all procedures with your facilities. Your would find useful information in BD website or http://www.cyto.purdue.edu/. And according to my personal experience, we failed to prepare qualified RNA from 100000 sorted cells for gene arrays. But the data is based on our FACS and microarray facility. May it help you and good luck.
Je pense, donc je suis




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