mRNA Differential Display PCR (mRNA DD-PCR)

mRNA Differential Display PCR is a laboratory technique which is used to compare and identify changes in gene expression at the mRNA level between two or more eukaryotic cell and tissue samples. In this assay, mRNA formed after transcription, which is further tranlated to the respective protein is reverse transcribed using a set of 3' "anchored oligopolydeoxythymidine primers" to generate cDNA library for each sample. cDNA fragment is further amplified using arbitrary 3' primers and anchored 3' primers (same as used for RT). The amplication products are subjected to electrophoresis.

Fig. 1 Experimental procedures of mRNA Differential Display PCR

• Extraction of total RNA
• Removal of DNA from total RNA
• Design of primers
• Reverse transcription of mRNA
• Selective PCR amplification of cDNA
• Gel Electrophoresis of PCR amplication products
• Gene sequencing

• Allow for the rapid and sensitive survey of gene expression using small amounts of mRNA.
• Allow for the simultaneous analysis of multiple samples.
• Able to identify the differential expression of genes with extremely low expression using PCR amplication technology.
• Able to visualize all the expressed genes in a eukaryotic cell in a systematic and sequence-dependent manner by using multiple primer combinations.

Thanks to its simplicity, sensitivity and reproducibility, the mRNA Differential Display PCR is widely used in developmental biology, cancer research, neuroscience, pathology, endocrinology, plant physiology, and many other fields.

Laminar Flow Hoods

Laminar Flow Hoods are mostly enclosed systems that use filters and directrional air flow to provide a contaminant-free work area for applications requiring a sterile work space, such as tissue culture, culture media preparation, and so on.

Benchtop Centrifuges

Centrifuges separate or concentrate substances suspended in a liquid medium by density. Space-saving fixed- and variable-speed benchtop or tabletop centrifuges are used for applications including tissue culture, protein work, DNA/RNA research, and cell harvesting.

Thermal Cycler

Thermal Cycler is a laboratory apparatus that amplify segments of DNA using PCR. A thermal cycler is also commonly called a DNA Amplifier, PCR Machine, or Thermocycler. The cycle normally lowers or raises the block’s temperature in preprogrammed discrete steps. A thermal cycler is important for a laboratory dealing in molecular biology and gene cloning.

Electrophoresis Systems

Electrophoresis equipment applies an electric charge to molecules, causing them to migrate towards their oppositely charged electrode. The technique is found in all research and clinical laboratories utilizing DNA and protein applications, and is divided into gel and capillary techniques. Choice of electrophoresis equipment depends on the molecule of study, appropriate method of separation and downstream application.

PCR Consumables

PCR consumables generally involve pipettes, 100ul, 500ul and 1ml centrifuge tubes, 12 consecutive rows of tubes, and tube racks (96-well plates, etc.) for placing the centrifuge tubes. It is best to perform PCR experiments on ice, so you need to prepare an ice box. We use a wide variety of PCR consumables for you to choose from to meet your different needs.

STEMart offers complete instruments and equipments used in mRNA Differential Display PCR to facilitate your research. If you want to learn more detail about the related devices, or would like to consult with the experts at STEMart, please feel free to contact us.

Reference

• Adam, Paul J. . "The Use of Differential mRNA Display (DDRT-PCR) to Identify Genes Differentially Expressed in Normal and Diseased Vascular Cells." Methods in Molecular Medicine 30(1999):99-110.