Identification of multiple mRNA and DNA sequences from small tissue samples isolated by laser-assisted microdissection.
Molecular analysis of small tissue samples has become increasingly important in biomedical studies. Using a laser dissection microscope and modified nucleic acid isolation protocols, we demonstrate that multiple mRNA as well as DNA sequences can be identified from a single-cell sample. In addition, we show that the specificity of procurement of tissue samples is not compromised by smear contamination resulting from scraping of the microtome knife during sectioning of lesions. The procedures described herein thus allow for efficient RT-PCR or PCR analysis of multiple nucleic acid sequences from small tissue samples obtained by laser-assisted microdissection.
Bernsen
M R
MR
Department of Pathology, University Hospital Nijmegen, The Netherlands.
Dijkman
H B
HB
de Vries
E
E
Figdor
C G
CG
Ruiter
D J
DJ
Adema
G J
GJ
van Muijen
G N
GN
D001483:Base Sequence; D018929:Cell Culture Techniques / Q000379:methods; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004210:Dissection / Q000379:methods; D006801:Humans; D019169:Jurkat Cells; D007834:Lasers; D008545:Melanoma / Q000235:genetics* / Q000473:pathology; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D012334:RNA, Neoplasm / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D004273:DNA, Neoplasm; D012333:RNA, Messenger; D012334:RNA, Neoplasm