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Methods Mol Biol. 2016;1379:201-11. doi: 10.1007/978-1-4939-3191-0_18.

South African Herbal Extracts as Potential Chemopreventive Agents: Screening for Anticancer Splicing Activity.

Author information

1
Research, Innovation & Engagements, Mangosuthu University of Technology, 392, Durban, 4031, South Africa. dlaminiz@mut.ac.za.
2
Mangosuthu University of Technology Research, Innovation and Engagements Portfolio, Durban, 4031, South Africa.
3
Faculty of Medicine and Health Sciences, Division of Pre-clinical Oncology, University of Nottingham, Nottingham, UK.

Abstract

RT-PCR is an invaluable tool for the detection and characterization of mRNA. Cancer cell lines are treated with crude plant extracts and RNA is extracted and purified with DNase prior to RT-PCR. RT-PCR first-strand cDNA synthesis is done using random primers and can be refrigerated at 4 °C. PCR from the stored cDNA is performed using transcript-specific primers and electrophoresed on a molecular grade agarose gel to separate the splice variants.

KEYWORDS:

Agarose gel electrophoresis; Anticancer splicing activity; DNase treatment; First-strand cDNA synthesis; Herbal plant extracts; RNA extraction

PMID:
26608302
DOI:
10.1007/978-1-4939-3191-0_18
[Indexed for MEDLINE]

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