Protocols for LCM preparation and analysis (NIH)

Original protocol from NIH laser capture microdissection core facility.

Content:

I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections
A. Embedding
B. Cutting
C. Staining
II. Preparation and LCM of Paraffin Embedded Tissue Sections
A. Fixation
B. Processing, Embedding and Tissue Sectioning
C. Staining


For both I & II Frozen & Paraffin-EmbeddedTissue Sections


D. Laser Transfer
E. DNA Extraction Protocol of LCM tissue and PCR for DNA analysis

F. RNA Extraction Protocol of LCM tissue and RT-PCR for mRNA analysis


Standard Protocols for Microdissected Tissue Analysis:
RNA Isolation (modified Stratagene Microisolation protocol)
DNAse
Re-extracton of RNA (same reagents as above)
Reverse Transcription (final volume 20 uL)
PCR
P.A.G.E. *(only if radioactivity is used)

Last update 18-Mar-2005, Rating n/a of 0 votes.