Recovering DNA and optimizing PCR conditions from microdissected formalin-fixed and paraffin-embedded materials.
Microdissection is a powerful technique in molecular pathology and genetic investigations. To detect genetic alterations such as gene mutation or deletion from tumor specimen, the purity of target cells is extremely critical. Unwanted cell contamination will dramatically dilute the detectable level of the abnormality. The major obstacle in clinical research is to obtain sufficient and qualified DNA from a small amount of formalin-fixed and paraffin-embedded materials. We have successfully modified our previous protocols and overcome the difficulties of recovery of DNA. After these modifications, almost every single formalin-fixed and paraffin-embedded specimen has been successfully amplified in the required DNA region.
Department of Genetics and Pathology, Rudbeck Laboratory, Unversity Hospital, Uppsala, Sweden. zhi-ping.ren@genpat.uu.se
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Ren
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Sällström
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Sundström
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Nistér
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Olsson
D004247:DNA / Q000302:isolation & purification*; D004210:Dissection / Q000379:methods*; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D016612:Paraffin Embedding / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods
D005404:Fixatives; D005557:Formaldehyde; D004247:DNA