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. Author manuscript; available in PMC: 2013 May 15.
Published in final edited form as: J Neurosci Methods. 2012 Mar 10;206(2):200–204. doi: 10.1016/j.jneumeth.2012.02.023

Fig. 2. Expression profiling of BMEC by immuno-LCM/TLDA with no RNA purification shows high reproducibility.

Fig. 2

Immuno-LCM of BMEC was carried out on separate EAE cerebellar sections, and captured tissue extracted with Cell Lysate Buffer® then processed through reverse transcription and cDNA pre-amplification, as in Fig. 1. To gauge sensitivity and exclusively highlight technical reproducibility, different dilutions (1/4 or 1/32) of pre-amplified cDNA derived from 500 (a) or 1000 (b) LCM shots were analyzed by qrt-PCR using the Immune Panel TLDA, and linear regression performed on the two corresponding raw Ct values obtained for each gene at the respective dilutions. To highlight overall reproducibility, while accounting for all potential variation due to both biological and technical components, the results of 500 LCM shots from two different tissue sections were compared (c), and linear regression performed on the two corresponding raw Ct values obtained for each gene from the respective sections.