ere extremely reproducible: macula-enriched target transcript
in the macula-derived samples was 19.9 0.2 cycles (n = 11), and 21.4
0.3 cycles (n = 11) in the periphery-derived samples (compare Figures
2, 3, and 4A). Here we have demonstrated that the use of iQ supermix to
optimize reaction conditions allows the best consistency and reproducibility
from experiment to experiment.
We are greatly indebted to Drs Gregory J Riggins and Kathy Boon, Duke
University Medical Center, Durham, NC, for their helpful expert advice.
Supported in part by NEI R01 EY 11286, NEI P30 EY 05722 and a Career Development
Award from Research to Prevent Blindness (CBR).
Boeckman F et al., Real-time PCR: General considerations, Bio-Rad bulletin
Bracete AM et al., Isolation of total RNA from small quantities of tissue
and cells, Focus 21, 3839 (1999)
Loging WT et al., Identifying potential tumor markers and antigens by
database mining and rapid expression screening, Genome Res 10, 13931402
Morrison TB et al., Quantification of low-copy transcripts by continuous
SYBR Green I monitoring during amplification, Biotechniques 24, 954962
The polymerase chain reaction (PCR) process is covered by patents owned
by Hoffman-LaRoche. Use of the PCR process requires a license.
SYBR is a trademark of Molecular Probes, Inc.
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