HOPE--a new fixing technique enables preservation and extraction of high molecular weight DNA and RNA of > 20 kb from paraffin-embedded tissues. Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect.
The growing number of molecular pathologic tools that are currently available require material with good long term preservation of morphology, nucleic acids, and antigenic structures. However, pathologic investigations of tissues done at a molecular level are often hampered by the fixatives in use. We thus endeavored to design a new fixing system, including subsequent paraffin-embedding and sectioning, that makes complete pathologic analyses possible, with special consideration of immunohistochemistry (IHC), in situ hybridization (ISH), and molecular pathology. The optimized HOPE (Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect) fixing technique allows us to preserve and extract high molecular weight DNA and RNA of > 20 kbp suitable for downstream applications, such as PCR and RT-PCR from HOPE-fixed, paraffin-embedded tissues that are up to 5 years old. This technique will most probably lead to new impacts on molecular pathology.
Wiedorn
Klaus Hermann
KH
Institut für Pathologie, Klinikum Stuttgart, Kriegsbergstr. 60, 70174 Stuttgart, Germany. k.wiedorn@katharinenhospital.de
Olert
Jürgen
J
Stacy
Robin A P
RA
Goldmann
Torsten
T
Kühl
Heike
H
Matthus
Jutta
J
Vollmer
Ekkehard
E
Bosse
Alexander
A
D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000032:analysis / Q000302:isolation & purification*; D005557:Formaldehyde; D006531:HEPES; D006801:Humans; D016612:Paraffin Embedding; D012313:RNA / Q000032:analysis / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016707:Tissue Fixation / Q000379:methods*
D003432:Cross-Linking Reagents; D005557:Formaldehyde; D012313:RNA; D004247:DNA; D006531:HEPES