PCR primers and probes used in the study:
Characterization of In Vitro and In vivo Human Neuroblastoma Models for the Study of Radioactive MIBG Uptake
by: Celso Castaldo, Elizabeth Melendez, Rex A. Moats, C. Patrick Reynolds
Previously published primer and probe sequences were employed for NAT mRNA expression analysis (GenBank accession number M65105) (15).
Primers and probes for the other genes were designed using a software program (Primer Express V1.5; Applied Biosystems) based on the sequences from GenBank/EMBL/DDBJ.
Stathmin 2 (NM_005419)
forward primer sequence: 5'-GCTGTCCATGCTGTCACTGATC-3'
reverse primer sequence: 5'-CTTCCATATCATCGTAAGTATAGATGTTGA-3'
Chromogranin A (NM_001275)
forward primer sequence: 5'-GGCACATCAGCAGAAGAAACAC-3'
reverse primer sequence: 5'-CTCTTTTCTCCATAACATCCTTGGA-3'
probe: 5'-TTCCACCGC CTGTTTCAGCTCGG-3'
Dopamine ß-hydroxylase (NM_000787)
forward primer sequence: 5'-AGATCGTGAACCAGGACAATCA-3'
reverse primer sequence: 5'-TCCGTGTTGTACGTGCAGGA-3'
Cyclin D1 (NM_053056)
forward primer sequence: 5'-CCGAGAAGCTGTGCATCTACAC-3'
reverse primer sequence: 5'-AGGTTCCACTTGAGCTTGTTCAC-3'
All the probes were labeled with the fluorescent reporter dye 6-carboxyfluorescein at the 5´ end and with the quencher molecule 6-carboxytetrametylrhodamine at the 3´ end.
The house-keeping gene GAPDH was used as an internal standard for all real-time PCR reactions.
Primers and probes were obtained from Integrated DNA Technologies (Coralville, Iowa, USA).