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Figure 2. Amplification of RNA from cell-free saliva for microarray study. (A) Monitoring of RNA amplification by agarose gel electrophoresis. Lanes 1 to 5 are 1-kb DNA ladder, 5 µL saliva after RNA isolation (undetectable), 1 µL of 2 round amplified cRNAs (range from 200 bp to ~ 4 kb), 1 µL cRNA after fragmentation (around 100 bp), and Ambion RNA Century Marker, respectively. (B) ACTB can be detected in every main step during salivary RNA amplification. The agarose gel shows an expected single band (153 bp) of PCR product. Lanes 1 to 8 are 100-bp DNA ladder, total RNA isolated from cell-free saliva, 1st round cDNA, 1st round cRNA after RT, 2nd round cDNA, 2nd round cRNA after RT, positive control (human total RNA, BD Biosciences Clontech, Palo Alto, CA, USA), and negative control (templates omitted), respectively. (C) Target cRNA analyzed by Agilent 2100 bioanalyzer before hybridization on microarray. Only one single peak in a narrow range (50–200 bp) was detected demonstrating proper fragmentation.
J DENT RES, Vol. 83, No. 3,
199-203 (2004)
DOI: 10.1177/154405910408300303
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