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H > ¼­ºñ½º > Next-Generation Sequencing (NGS) > RNA-Seq(Transcriptome)

RNA-Seq(Transcriptome)

Prokaryotes RNA-Seq (Transcriptome)

Prokayotes RNA-Seq(Transcriptome)Àº ½ÇÇè ´ë»ó ¹Ì»ý¹°Ã¼³»¿¡¼­ ¹ßÇöµÇ´Â ¸ðµç transcript (mRNA, non-coding RNA Æ÷ÇÔ)¿¡ ´ëÇÑ ¹ßÇö Á¤·®È­ (quantification) ¹× ±¸Á¶ ºÐ¼® (strucutural analysis)À» °á°ú¹°·Î ¾òÀ» ¼ö ÀÖ´Â NGS ±â¹ýÀÔ´Ï´Ù. Prokaryotes RNA-Seq(Transcriptome) ¹æ¹ýÀ» ÀÌ¿ëÇÏ¿©, ÇØ´ç ¹Ì»ý¹°Ã¼³»ÀÇ transcriptome¿¡ ´ëÇÑ ´ëºÎºÐÀÇ Á¤º¸¸¦ ¾òÀ¸½Ç ¼ö ÀÖÀ¸¸ç, À̸¦ ÀÌ¿ëÇÏ¿© ±âÃÊ ºÐ¾ß »Ó ¾Æ´Ï¶ó ÀÇÇÐ ¹× ¾à¹° À¯ÀüüÇÐ µî ´Ù¾çÇÑ ºÐ¾ß¿¡ Àû¿ëµÉ ¼ö ÀÖ½À´Ï´Ù.

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  • Transcriptome ½ÃÄö½Ì (>95% coverage)
  • novel coding / non-coding RNA ¹ß±¼
  • gene structural analysis (alternative splicing, fusion genes)
  • transcriptome ³»ÀÇ SNP ºÐ¼®
  • differential gene expression analysis

½ÇÇè Workflow

Bioinformatics ºÐ¼® Workflow

RNA-Seq(Transcriptome)-½ÇÇè ÁøÇà °úÁ¤

Sample requirements

concentration Concentration (ng/§¡) Quality
Total RNA > 2§¶ (human, mouse, rat)
> 5§¶ (other animals)
> 80 (human, mouse, rat)
> 200 (other animals)
OD(260/230) > 2.0, OD(260/280)>1.8 is highly recommended

Sequencing Strategy

50 SE sequencing

Bioinformatic analysis - contents

RNA-Seq(Transcriptome)-resequencing
1Â÷ ºÐ¼® 1. Data filtering includes removing adaptors, contamination and low-quality reads from raw reads
2. Assessment of sequencing£¨Statistics of raw reads, Sequencing saturation analysis,
    Analysis of the distribution of reads on reference gene£©
3. Gene expression annotation
2Â÷ ºÐ¼®

4. Differential gene expression analysis (Screening of differentially expressed genes(DEGs), Experimental repeatability analysis of DEGs.

5. Expression pattern analysis of DEGs (Client to determine what conditions to meet to perform cluster analysis, for example, "hormone treatment in 2 hours, 4 hours, 8 hours showed a significant difference of gene expression")

6. Gene ontology analysis of DEGs

7. Pathway enrichment analysis of DEGs

8. Protein-protein interaction network analysis (protein-protein interaction database of the target species needed)

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