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Revista Colombiana de Biotecnología

Print version ISSN 0123-3475

Abstract

ZABALETA VANEGAS, Milet; YAYA LANCHEROS, Mary Luz  and  CHAPARRO GIRALDO, Alejandro. Comparison of two kits of RT-PCR in RNAm Detection of two endegenous genes of potato (Solanum tuberosum spp. Andigena). Rev. colomb. biotecnol [online]. 2008, vol.10, n.2, pp.104-110. ISSN 0123-3475.

RT-PCR is a technique which using a RNAm like template is obtained a cDNA (complementary DNA) for reverse transcription, then is amplified one of them by PCR using specifics primers. This technique allows make expression studies to mRNA level. In order to implement this technique in potato (Solanum tuberosum spp. Andigena), was used in vitri grow plants Pastusa Suprema variety. Initially was established the total RNA extraction conditions using the Invitrogen ™ TRizol® Reagent kit., with this the results were successful. Extracted RNA was used as a template to probe two Invitrogen™ kits: "ONE Step superScript ™"and "SuperScript™ First Strand Syntesis For RT-PCR SS™ II RT". The specifics primers were designed for endogenous genes: cox and actin. The first one is a mitocondrial gene and the second one is a nuclear gene. Defined amplification signals for cox, using "ONE Step superScript ™" kit, whit a spected 96 bp size. In actin, a clear amplification signal was observed (size 300 bp) with the "SuperScript™ First Strand Syntesis For RT-PCR SS™ II RT" kit.

Keywords : Solanum tuberosum; cox; actin; RT-PCR; potato.

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