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Laser Capture Microdissection | Troubleshooting Forum

10/09/2012
Kristie Nybo, Ph.D.

What can I do to improve the yield and quality of the RNA?


The BioTechniques Molecular Biology Forum brings bench scientists together in an online community to ask for and share advice regarding laboratory problems. This week’s highlighted question comes from the Microscopy and Imaging Techniques forum.

I am trying to use laser capture microdissection to collect cells from paraformaldehyde-fixed midbrain sections from transgenic mice. What can I do to improve the yield and quality of the RNA? Source: NIH.gov





Q: I am trying to use laser capture microdissection to collect cells from paraformaldehyde-fixed midbrain sections from transgenic mice. The quantity and quality of RNA isolated were insufficient for real-time PCR. I don’t need to stain the tissue because the cells of interest express GFP, but because of this, I need to avoid alcohol dehydration. What can I do to improve the yield and quality of the RNA?

A: For isolating RNA, you need to use a membrane slide if you are using a 2 laser system. I would recommend that the samples…

A: Remember not to rely on oligo dT primers for the reverse transcription step if you get enough RNA out of your tissue because formaldehyde modifies the adenine in poly-A tails and therefore…

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