Gene Expression
Nebulas
Gene Expression NebulasSummary: Sorghum is an important cereal crop, which requires large quantities of nitrogen fertilizer for achieving commercial yields. Identification of the genes responsible for low-N tolerance in sorghum will facilitate understanding of the molecular mechanisms of low-N tolerance, and also facilitate the genetic improvement of sorghum through marker-assisted selection or gene transformation. In this study we compared the transcriptomes of root tissues from seven sorghum genotypes having different genetic backgrounds with contrasting low-N tolerance by the RNAseq deep sequencing data. Several genes were found which are common differentially expressed genes between four low-N tolerant sorghum genotypes (San Chi San, China17, KS78 and high-NUE bulk) and three sensitive genotypes (CK60, BTx623 and low-NUE bulk).
Overall Design: RNAseq deep sequencing
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| Growth Protocol: | The seeds were grown in three inch pots under a 16/8 h photoperiod at 25°C (day) and 18°C (night). |
| Treatment Protocol: | No treatment |
| Extract Protocol: | Total RNA was extracted first using NTES buffer (20 mM TRIS pH 8, 10 mM EDTA, 100 mM NaCl and 1% SDS) and followed by Trizol reagent (Invitrogen) using the manufacturer’s instructions. |
| Library Construction Protocol: | RNA libraries were prepared from 4 μg total RNA using the Illumina TruSeq RNA Sample Prep Kit v2 - Set A (RS-122-2002) according to the manufacturer’s instructions. Libraries were analyzed and measured by gel electrophoresis and NanoDrop 1000 Spectrophotometer to a concentration of 10 nM each. Four indexed libraries were pooled into one lane and clusters generated at 8 pM concentration were sequenced on the Illumina Genome Analyzer IIx (GAIIx; Illumina, Inc., San Diego, CA) using three 36-cycle sequencing kits to read 76 nucleotides of sequence from a single end of each insert, by standard multiplexing v8.3 protocol. |
| Molecule Type: | poly(A)+ RNA |
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| Library Layout: | SINGLE |
| Library Strand: | - |
| Platform: | ILLUMINA |
| Instrument Model: | Illumina Genome Analyzer IIx |
| Strand-Specific: | Unspecific |