Abstract
Reverse-transcription polymerase chain reaction (RT-PCR) was used to generate a panel of distinct amplicons from total RNA derived from a variety of sources. Following amplification, semi-quantification of the different amplicons was performed by normalization to steady-state transcript levels of the house-keeping gene, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), using laser densitometry, phosphorimaging, and liquid scintillation counting. In this report we demonstrate that incorporation of [α-32P] dCTP into the PCR amplicons followed by band excision from agarose gels, and counting by liquid-scintillation is both cost-effective and reproducible method for mRNA transcript semi-quantitation. © Rapid Science Ltd. 1998
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Sharma, V., Xu, M., Vail, J. et al. Comparative analysis of multiple techniques for semi-quantitation of RT-PCR amplicons. Biotechnology Techniques 12, 521–524 (1998). https://doi.org/10.1023/A:1008847329652
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DOI: https://doi.org/10.1023/A:1008847329652