Quantitative Gene Expression

Gene expression refers to the process in which cells convert genetic information stored in DNA sequences into biologically active protein molecules in the process of life to perform various functions. Genes that are actively expressed generally account for about 10-15% of the genes contained in the genome, and researchers even overexpress or suppress the target fragments. Analyzing the expression of these genes has become an important part of life science research. Creative Biomart Biomarker provides you with different technical means, such as fluorescence quantitative PCR and transcriptome sequencing (RNA-seq), etc., which can study the RNA in various samples under various conditions and the changes of DNA related to expression regulation at different levels.

Gene expression detection can be divided into protein level expression detection and mRNA expression level detection.

Protein level expression detection can be divided into Western Blot, ELISA and HPLC-based quantitative detection analysis and protein function analysis. Protein function analysis can be divided into the following parts:

  • Protein subcellular localization analysis: immunofluorescence technology;
  • Protein interaction study: yeast two-hybrid, immunoprecipitation, fluorescence resonance energy transfer (FRET)
  • Enzyme activity detection: ELISA, HPLC

Detection of mRNA expression levels can be divided into:

  • Semi-quantitative RT-PCR
  • Northern Blot
  • Real-time quantitative PCR.

Creative Biomart Biomarker can provide services according to your different needs. You can click the service name to display the corresponding page. We introduce you Northern Blot and qRT- PCR as examples:

Northern Blot

The experimental method used to detect the expression level and size of eukaryotic RNA to estimate its abundance can obtain data simultaneously through a large number of experimental samples.

The Protocol of Northern Blot:

  1. Separation of complete mRNA
  2. Separation of RNA by agarose gel
  3. Transfer film
  4. UV cross-linking (fixing RNA to the support)
  5. Solid phase molecules hybridize with DNA or RNA molecules and remove non-specific hybridized probe molecules
  6. Detection, capture and analysis of specifically bound RNA molecules

Fluorescence quantitative PCR (dye method or probe method)

Add a fluorescent group to the PCR system, use the change of the fluorescent signal to detect the change of each amplification product in the PCR amplification reaction in real time, and quantitatively analyze the starting template through the relationship between the Ct value and the standard curve or internal reference gene.

The Protocol of qRT-PCR:

  1. Target gene analysis and primer design (including dye method or probe method selection)
  2. RNA or microRNA extraction
  3. Reverse transcription experiment
  4. Real-time PCR
  5. Analysis of relative expression or absolute expression

We can provide services not limited to the following:

  1. Conventional fluorescence quantitative PCR:
  2. Gene expression analysis of animal samples, plant samples, bacteria and fungi samples; expression profile chip, transcriptome sequencing results verification; RNAi interference target gene test results

  3. MicroRNA fluorescence quantitative PCR:
  4. Verification of microRNA chip and microRNA sequencing experiment results

  5. Noncoding RNA fluorescence quantitative PCR:
  6. Analysis of the expression of non-coding RNA of various lengths in various tissues of humans and mice

  7. Other applications:
  8. Analysis of copy number changes of specific genes in the genome, or absolute quantification of the total number of microorganisms; qualitative detection of viruses and pathogens, qualitative identification of biological species.

    At the same time, we provide accurate quantitative gene expression services. Only a specific lysate is required to lyse the sample, and specific particles can specifically recognize the target mRNA. The signal intensity is proportional to the amount of mRNA, and the gene can be accurately quantified by detecting the fluorescence intensity.

The Quantitative Gene Expression service provided by Creative Biomart Biomarker has the following advantages:

  • Strict quality control in each link, including primer design, optimization of PCR conditions, specific melting curve, template quality, etc.
  • The target gene and internal reference gene are both international standard 3 multi-well experiments, and the Ct value of the multi-well is guaranteed to be uniform
  • For the accurate quantification service, we have no requirements on the lower limit of the sample, and can perform multiple gene accurate quantification in the same reaction

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