RNA extraction and primary screening of reference genes in Camellia oleifera
Author(s): SONG Zhi-bo, LIU Min, JIA Bao-guang, TIAN Lei, ZENG Yan-ling, ZHOU Jun-qin, TAN Xiao-feng, ZHANG Lin
Pages: 93-98
Year: 2014 Issue: 2
Journal: Nonwood Forest Research
Keyword: Camellia oleifera; RNA extraction; reference gene; reverse transcription PCR;
Abstract: In order to obtain high quality total RNA and stable reference genes in Camellia oleifera, taking ifne cultivar‘Huashuo’ as materials, total RNA was extracted by a combination of ambion kit and CTAB method. The obtained total RNA has good integrity and high purity. A total of 14 genes were selected as candidates of reference genes based on the previously constructed transcriptome data in C. oleifera, and 22 primers were designed for the 14 genes. The reference genes suitable for different tissues or organs were rapidly screened out by RT-PCR reaction system optimization and agarose gel electrophoresis detection. ALB (albumin gene), EF1α (elongation factor gene), ETIF3H (eukaryotic translation initiation gene), UBC2 (ubiquitin-conjugating enzyme gene) can be used as the reference genes for fruits, EF1αcan be used as a reference gene for root, stem, lfower and fruit, and ETIF3H can be used as a reliable reference gene for root, stem, leaf and fruit in C. oleifera.
Pages: 93-98
Year: 2014 Issue: 2
Journal: Nonwood Forest Research
Keyword: Camellia oleifera; RNA extraction; reference gene; reverse transcription PCR;
Abstract: In order to obtain high quality total RNA and stable reference genes in Camellia oleifera, taking ifne cultivar‘Huashuo’ as materials, total RNA was extracted by a combination of ambion kit and CTAB method. The obtained total RNA has good integrity and high purity. A total of 14 genes were selected as candidates of reference genes based on the previously constructed transcriptome data in C. oleifera, and 22 primers were designed for the 14 genes. The reference genes suitable for different tissues or organs were rapidly screened out by RT-PCR reaction system optimization and agarose gel electrophoresis detection. ALB (albumin gene), EF1α (elongation factor gene), ETIF3H (eukaryotic translation initiation gene), UBC2 (ubiquitin-conjugating enzyme gene) can be used as the reference genes for fruits, EF1αcan be used as a reference gene for root, stem, lfower and fruit, and ETIF3H can be used as a reliable reference gene for root, stem, leaf and fruit in C. oleifera.
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