RNA-Seq (Quantification)
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RNA-Seq (Quantification) is used to analyze gene expression of certain biological objects under specific conditions. It is a cost-effective quantification method that produces greater sensitivity, reproducibility, and wider dynamic range than microarrays. BGI quantifies whole-genome gene expressions using next-gen sequencer Illumina HiSeq 2000 and provides differential gene expression and pathway analysis. Powered by our unique bioinformatics capabilities, BGI’s RNA-Seq services deliver reliable data and analyses to our customers conducting drug response studies, biomarker detection, basic medical research, and drug R&D.

Benefits:

Compared to DNA microarray, BGI’s RNA-Seq (Quantification) service offers:

  1. Greater reproducibility
  2. Greater specificity - more accurate, no cross-hybridization, no restrictions to the available probes
  3. Greater sensitivity- detects 50% more genes, wider dynamic range.
  4. Strong bioinformatics analysis capabilities on RNA-Seq data
  5. Results compatible with common analysis software

BGI has validated the reproducibility, sensitivity, and detection range of RNA-Seq(Quantification).
Some results are as follows:

Materials: Human Brain Reference RNA (HBRR) from FirstChoice ®;

Universal Human Reference RNA (UHRR) from Stratagene

Standardized method of gene expression analysis: RPKM (Reads per kb per million reads) is used to quantify the gene expression level. The method is able to eliminate the influence of different gene lengths and sequencing discrepancy in the measurement.

High Reproducibility

RNA Seq
Figure 1: Technical Replicate Results of RNA-Seq (Quantification). From the technical replicate data, the spearman R is 0.992 and 0.993, respectively.

The results suggest that RNA-Seq has high reproducibility.

High Accuracy

RNA-Seq_HighAccuracy
Figure 2: Correlation between results from RNA-Seq (Quantification) and qPCR RNA-Seq is a highly accurate quantitation method. The scatter-plot of figure 2 show a high correlation between results from RNA-Seq and qPCR. Spearman R is 0.915 and slope is 1.022.

High Sensitivity

RNA-Seq_HighSensitivity
Figure 3: Detection Range of RNA-Seq (Quantification) is wider than microarray, and RNA-Seq can detect more low abundance genes than micriarray.

Bioinformatics:

  1. Differentially expressed genes analysis
  2. Expression pattern analysis
  3. GO functional enrichment analysis
  4. Pathway enrichment analysis
  5. Protein-protein interaction network analysis

Sample Requirements:

For the total RNA samples you provide us:

  1. Purity: OD260/280=1.8-2.2; OD260/230 ≥ 2.0, RIN≥7.0; 28S:18S>1.0
  2. Plants Samples: no less than 20ug total RNA for library preparation, and concentration≥400ng/μl.
  3. Other Eukaryotes: more than 5μg total RNA is necessary, and concentration ≥200 ng/μl.
  4. For Human and Mouse Samples: Starting at 5μg total RNA. Concentration≥80 ng/μl is recommended.


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