TY - JOUR
T1 - The normalization of gene expression data in melanoma
T2 - Investigating the use of glyceraldehyde 3-phosphate dehydrogenase and 18S ribosomal RNA as internal reference genes for quantitative real-time PCR
AU - Giricz, Orsolya
AU - Lauer-Fields, Janelle L.
AU - Fields, Gregg B.
N1 - Funding Information:
We gratefully acknowledge support of this work from the National Institutes of Health (CA 98799 and EB 00289 to G.B.F.) and the FAU Center of Excellence in Biomedical and Marine Biotechnology.
PY - 2008/9/1
Y1 - 2008/9/1
N2 - We examined a panel of 26 melanoma and fibroblast samples (tissues and cultured cells) to evaluate the suitability of two commonly used housekeeping genes, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and 18S ribosomal RNA (rRNA), for quantitative real-time PCR. Both genes showed significant variations within the individual cell line and tissue groups. Although no overall trends were observed in the expression of the 18S rRNA, GAPDH was up-regulated in melanoma tissue and cultured cells compared with the corresponding normal samples. In melanoma and fibroblast cell lines and tissues, absolute quantification appears to be more appropriate than normalizing messenger RNA (mRNA) expression via GAPDH or 18S rRNA housekeeping genes.
AB - We examined a panel of 26 melanoma and fibroblast samples (tissues and cultured cells) to evaluate the suitability of two commonly used housekeeping genes, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and 18S ribosomal RNA (rRNA), for quantitative real-time PCR. Both genes showed significant variations within the individual cell line and tissue groups. Although no overall trends were observed in the expression of the 18S rRNA, GAPDH was up-regulated in melanoma tissue and cultured cells compared with the corresponding normal samples. In melanoma and fibroblast cell lines and tissues, absolute quantification appears to be more appropriate than normalizing messenger RNA (mRNA) expression via GAPDH or 18S rRNA housekeeping genes.
UR - http://www.scopus.com/inward/record.url?scp=46649089701&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=46649089701&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2008.05.024
DO - 10.1016/j.ab.2008.05.024
M3 - Article
C2 - 18554498
AN - SCOPUS:46649089701
SN - 0003-2697
VL - 380
SP - 137
EP - 139
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -