A Modified Hot Borate Method Significantly Enhances the Yield of High-Quality RNA from Cotton (Gossypium hirsutum L.)

Wan, Ching-Yi; Wilkins, Thea A.

doi:10.1006/abio.1994.1538

Cited by 912 publications

(513 citation statements)

References 0 publications

Supporting

Mentioning

511

Contrasting

Unclassified

Order By: Relevance

“…RNA from hormone-treated plant materials was prepared using the Invisorb Spin Plant RNA mini kit (Invitek GmbH), according to the manufacturer's protocol. RNA samples from green siliques and seeds were prepared using a ''hot borate'' method (Wan and Wilkins, 1994).…”

Section: Rna Extractionmentioning

confidence: 99%

Genome-Wide Identification and Testing of Superior Reference Genes for Transcript Normalization in Arabidopsis

et al. 2005

View full text Add to dashboard Cite

Gene transcripts with invariant abundance during development and in the face of environmental stimuli are essential reference points for accurate gene expression analyses, such as RNA gel-blot analysis or quantitative reverse transcription-polymerase chain reaction (PCR). An exceptionally large set of data from Affymetrix ATH1 whole-genome GeneChip studies provided the means to identify a new generation of reference genes with very stable expression levels in the model plant species Arabidopsis (Arabidopsis thaliana). Hundreds of Arabidopsis genes were found that outperform traditional reference genes in terms of expression stability throughout development and under a range of environmental conditions. Most of these were expressed at much lower levels than traditional reference genes, making them very suitable for normalization of gene expression over a wide range of transcript levels. Specific and efficient primers were developed for 22 genes and tested on a diverse set of 20 cDNA samples. Quantitative reverse transcription-PCR confirmed superior expression stability and lower absolute expression levels for many of these genes, including genes encoding a protein phosphatase 2A subunit, a coatomer subunit, and an ubiquitin-conjugating enzyme. The developed PCR primers or hybridization probes for the novel reference genes will enable better normalization and quantification of transcript levels in Arabidopsis in the future.

show abstract

Section: Rna Extractionmentioning

confidence: 99%

Genome-Wide Identification and Testing of Superior Reference Genes for Transcript Normalization in Arabidopsis

et al. 2005

View full text Add to dashboard Cite

show abstract

“…Total RNA was isolated from 1 g apple tissue by a hot borate method (Wan and Wilkins, 1994). Genomic DNA was isolated from 20 mg of young leaf material using a DNeasy plant mini kit (QIAGEN).…”

Section: Preparation Of Nucleic Acids and Cdna Synthesismentioning

confidence: 99%

Light-Induced Expression of a MYB Gene Regulates Anthocyanin Biosynthesis in Red Apples

Takos¹,

Jaffé²,

Jacob³

et al. 2006

Plant Physiol.

911

776

View full text Add to dashboard Cite

Anthocyanins are secondary metabolites found in higher plants that contribute to the colors of flowers and fruits. In apples (Malus domestica Borkh.), several steps of the anthocyanin pathway are coordinately regulated, suggesting control by common transcription factors. A gene encoding an R2R3 MYB transcription factor was isolated from apple (cv Cripps' Pink) and designated MdMYB1. Analysis of the deduced amino acid sequence suggests that this gene encodes an ortholog of anthocyanin regulators in other plants. The expression of MdMYB1 in both Arabidopsis (Arabidopsis thaliana) plants and cultured grape cells induced the ectopic synthesis of anthocyanin. In the grape (Vitis vinifera) cells MdMYB1 stimulated transcription from the promoters of two apple genes encoding anthocyanin biosynthetic enzymes. In ripening apple fruit the transcription of MdMYB1 was correlated with anthocyanin synthesis in red skin sectors of fruit. When dark-grown fruit were exposed to sunlight, MdMYB1 transcript levels increased over several days, correlating with anthocyanin synthesis in the skin. MdMYB1 gene transcripts were more abundant in red skin apple cultivars compared to non-red skin cultivars. Several polymorphisms were identified in the promoter of MdMYB1. A derived cleaved amplified polymorphic sequence marker designed to one of these polymorphisms segregated with the inheritance of skin color in progeny from a cross of an unnamed red skin selection (a sibling of Cripps' Pink) and the non-red skin cultivar Golden Delicious. We conclude that MdMYB1 coordinately regulates genes in the anthocyanin pathway and the expression level of this regulator is the genetic basis for apple skin color.

show abstract

“…RNA was extracted by the hot borate method (Wan and Wilkins, 1994). Poly(A) ϩ RNA was isolated using OligotexdT30 (Takara, Kyoto, Japan) according to the manufacturer's protocol.…”

Section: Rna Extraction and Rt-pcrmentioning

confidence: 99%

Differential Expression and Internal Feedback Regulation of 1-Aminocyclopropane-1-Carboxylate Synthase, 1-Aminocyclopropane-1-Carboxylate Oxidase, and Ethylene Receptor Genes in Tomato Fruit during Development and Ripening

et al. 1998

View full text Add to dashboard Cite

We investigated the feedback regulation of ethylene biosynthesis in tomato (Lycopersicon esculentum) fruit with respect to the transition from system 1 to system 2 ethylene production. The abundance of LE-ACS2, LE-ACS4, and NR mRNAs increased in the ripening fruit concomitant with a burst in ethylene production. These increases in mRNAs with ripening were prevented to a large extent by treatment with 1-methylcyclopropene (MCP), an ethylene action inhibitor. Transcripts for the LE-ACS6 gene, which accumulated in preclimacteric fruit but not in untreated ripening fruit, did accumulate in ripening fruit treated with MCP. Treatment of young fruit with propylene prevented the accumulation of transcripts for this gene. LE-ACS1A, LE-ACS3, and TAE1 genes were expressed constitutively in the fruit throughout development and ripening irrespective of whether the fruit was treated with MCP or propylene. The transcripts for LE-ACO1 and LE-ACO4 genes already existed in preclimacteric fruit and increased greatly when ripening commenced. These increases in LE-ACO mRNA with ripening were also prevented by treatment with MCP. The results suggest that in tomato fruit the preclimacteric system 1 ethylene is possibly mediated via constitutively expressed LE-ACS1A and LE-ACS3 and negatively feedback-regulated LE-ACS6 genes with preexisting LE-ACO1 and LE-ACO4 mRNAs. At the onset of the climacteric stage, it shifts to system 2 ethylene, with a large accumulation of LE-ACS2, LE-ACS4, LE-ACO1, and LE-ACO4 mRNAs as a result of a positive feedback regulation. This transition from system 1 to system 2 ethylene production might be related to the accumulated level of NR mRNA.

show abstract

A Modified Hot Borate Method Significantly Enhances the Yield of High-Quality RNA from Cotton (Gossypium hirsutum L.)

Cited by 912 publications

References 0 publications

Genome-Wide Identification and Testing of Superior Reference Genes for Transcript Normalization in Arabidopsis

Genome-Wide Identification and Testing of Superior Reference Genes for Transcript Normalization in Arabidopsis

Light-Induced Expression of a MYB Gene Regulates Anthocyanin Biosynthesis in Red Apples

Differential Expression and Internal Feedback Regulation of 1-Aminocyclopropane-1-Carboxylate Synthase, 1-Aminocyclopropane-1-Carboxylate Oxidase, and Ethylene Receptor Genes in Tomato Fruit during Development and Ripening

Contact Info

Product

Resources

About