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An improved method for RNA extraction from common bean seeds and validation of reference genes for qPCR
- Source :
- Crop Breeding and Applied Biotechnology, Volume: 17, Issue: 2, Pages: 150-158, Published: JUN 2017, Crop Breeding and Applied Biotechnology, Vol 17, Iss 2, Pp 150-158 (2017), Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA-Alice), Empresa Brasileira de Pesquisa Agropecuária (Embrapa), instacron:EMBRAPA, Crop Breeding and Applied Biotechnology v.17 n.2 2017, Crop Breeding and Applied Biotechnology, Sociedade Brasileira de Melhoramento de Plantas, instacron:CBAB
- Publication Year :
- 2017
- Publisher :
- Crop Breeding and Applied Biotechnology, 2017.
-
Abstract
- An RNA extraction method with high integrity and purity as well as the selection of adequate reference genes are prerequisites for gene expression analysis. For common bean seeds, there is no well-defined protocol that can be used in a laboratory routine for gene expression analysis. In this study, an extraction protocol for RNA from common bean seeds, which produced material with good integrity for qPCR (RIN ≥ 6.5), was optimized. In addition, 10 reference genes were evaluated under qPCR standard conditions using different tissue samples of common beans. Gene stabilities were analyzed using the delta-CT method, Bestkeeper, NormFinder and geNorm approaches. The genes β-tubulin and T197 were ranked as the most stable among the sample sets evaluated with different tissue samples, while PvAct and Pv18S were the least stable. To our knowledge, this is the first study evaluating RNA isolation methods and reference gene selection for seeds of Phaseolus vulgaris. Made available in DSpace on 2017-09-27T10:32:00Z (GMT). No. of bitstreams: 1 CNPAF2017cbab.pdf: 460872 bytes, checksum: b7233be3c24ab77be0d94d7156f8aac4 (MD5) Previous issue date: 2017-09-26
- Subjects :
- 0301 basic medicine
Beans
lcsh:Biotechnology
Improved method
Computational biology
lcsh:Plant culture
Phaseolus vulgaris
Gene normalization
Grain RNA extraction
03 medical and health sciences
Melhoramento
lcsh:TP248.13-248.65
Reference genes
Gene expression
lcsh:SB1-1110
Gene
General Environmental Science
biology
business.industry
Extraction (chemistry)
RNA
food and beverages
Molecular Breeding
biology.organism_classification
Biotechnology
030104 developmental biology
General Earth and Planetary Sciences
RNA extraction
Phaseolus
business
Feijão
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Crop Breeding and Applied Biotechnology, Volume: 17, Issue: 2, Pages: 150-158, Published: JUN 2017, Crop Breeding and Applied Biotechnology, Vol 17, Iss 2, Pp 150-158 (2017), Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA-Alice), Empresa Brasileira de Pesquisa Agropecuária (Embrapa), instacron:EMBRAPA, Crop Breeding and Applied Biotechnology v.17 n.2 2017, Crop Breeding and Applied Biotechnology, Sociedade Brasileira de Melhoramento de Plantas, instacron:CBAB
- Accession number :
- edsair.doi.dedup.....c6d494bd1a57f718e1330aa07aee68f1