转录组/Eukaryotic transcriptome sequencing

Macrogenome sequencing

Technical introduction

     A transcriptional group is the sum of all the transcripts of a specific tissue or cell in a functional state, including mRNA and non coded RNA (i.e., Non-coding RNA, such as tRNA, rRNAs, microRNAs, piRNAs, and long ncRNAs, etc.). Transcriptome sequencing (transcriptional sequencing) mainly refers to the study of mRNA which is transcribed by specific tissues or cells at a certain period through high throughput sequencing. It can be used to study gene structure and gene quantitative analysis and predict new transcripts. It is a powerful tool and tool to study the complexity of the transcriptional group. At present, transcriptome sequencing has been widely used in the study of animal and plant development, screening of good genes and disease treatment.

 

technology roadmap

 

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Suggestions for sending samples

物种浓度 (Qubit)体积总量DNA 完整性
真菌 total RNA
≥2 0ng/μL15 μL-100 μL≥2 μg

RIN≥6.5 28S/18S≥1.0;

图谱基线无上抬,5S 峰正常

其他动、植物

total RNA

≥20 ng/μL1 5μL-100 μL≥2 μg

RIN≥6.5 28S/18S≥1.0;

图谱基线无上抬,5S 峰正常
原核 total RNA≥35 ng/μL15 μL-100 μL≥2 μg

RIN≥6.5 23/18S≥1.0;

图谱基线无上抬,5S 峰正常

 

technical parameter

有参转录组测序测序策略数据量周期

HiSeq4000 PE150

6G clean date
45 个自然日

 

case analysis

Research background         

Variable shear plays an important role in plant growth and stress response, and there are many corresponding studies, but the interaction between growth and stress response is not very thorough.

research objective

The interaction between plant growth and stress reaction was explored by the analysis of the variable shear and gene expression changes in the seeds, germ and endosperm of the normal and arid groups of Maize by the transcriptional sequence.

Research results

48000 shear forms have been found in known genes, in which variable shear forms associated with development are affected by drought stress, and most of them occur in leaves and spikes, while male spikes are not affected. In the study of the correlation between the changes in the expression of variable shear related genes and the shear events of the corresponding genes, the gene expression of the shear factors in the development stages of the normal group was linearly related to the shear time, but in the drought group, the correlation was poor. These new findings again indicate the important role of variable shear in drought stress and growth and development of maize.

 

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Fig. 1. variable shear related gene expression and variable shear events

A map: expression correlation of variable shear related genes in various tissues.

B diagram: variable shear mode correlation in various tissues.

Reference

[1]. Thatcher SR, et al.  Genome-Wide Analysis of Alternative Splicing during Development and Drought Stress in Maize. Plant Physiol,(2015). 

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