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Working With Laser Capture Microdissection Samples

s of RNA recovered from LCM sections after various steps of the conventional post-sectioning processing protocols for frozen samples. These studies revealed that significant RNA degradation occurred during exposure of the section to aqueous solutions, presumably due to reactivation of endogenous nucleases. To circumvent this problem, Ambion developed a staining procedure that avoids exposing the sections to water at any step. (See our LCM Staining Kit.) Using the optimized "water-free" tissue processing protocol in conjunction with the RNAqueous-Micro Kit, total RNA of excellent quality, as assessed by microfluidic capillary electrophoresis and real-time RT-PCR, can be obtained from pure populations of cells selected by LCM (Figure 1).

Figure 1. Real-Time RT-PCR of ICM Samples. Mouse spleen (A) and liver (B) samples (1 mg, 3 mg and 10 mg) were dissected by LCM and RNA was isolated using the RNAqueous-Micro Kit according to protocol. The RNA was then used as a template for the amplification of GAPDH by real-time RT-PCR.


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Cat# Product Name Size 1931 RNAqueous-Micro Kit 50 purifications 1935 LCM Staining Kit Reagents for processing
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