title: Elimination of amplification artifacts in real-time reverse transcription PCR using laser capture microdissected samples.
abstract: Gene expression analysis on laser capture microdissected samples can be hampered because of the small sample size. The interference of PCR inhibitors increases with smaller sample size. Real-time reverse transcription PCR (RT-PCR) is usually performed directly after the reverse transcription step, enabling PCR inhibitors to remain in the complementary DNA (cDNA). A protocol was optimized for real-time RT-PCR with SYBR Green I. The introduction of an additional cDNA purification step after reverse transcription removed PCR inhibitors, making the reaction more efficient.
authors.affiliation: Department of Morphology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium. ward.despiegelaere@ugent.be
authors.forename: Ward
authors.initials: W
authors.lastname: De Spiegelaere
authors.forename: Tim
authors.initials: T
authors.lastname: Erkens
authors.forename: Jurgen
authors.initials: J
authors.lastname: De Craene
authors.forename: Christian
authors.initials: C
authors.lastname: Burvenich
authors.forename: Luc
authors.initials: L
authors.lastname: Peelman
authors.forename: Wim
authors.initials: W
authors.lastname: Van den Broeck
mesh_terms: D016477:Artifacts; D052160:Benzothiazoles; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003959:Diamines; D005456:Fluorescent Dyes / Q000378:metabolism; D007834:Lasers; D042282:Microdissection; D009930:Organic Chemicals / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D011804:Quinolines; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D013997:Time Factors
chemical_list: D052160:Benzothiazoles; D018076:DNA, Complementary; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D012313:RNA