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Analysis of EMSA by Real-Time PCR Method (CAT#: STEM-MB-2879-LGZ)

Introduction

A real-time polymerase chain reaction (real-time PCR,or qPCR) is a laboratory technique of molecular biology based on the polymerase chain reaction(PCR). It monitors the amplification of a targeted DNA molecule during the PCR(i.e.,in real time),not at its end,as in conventional PCR.<br />Two common methods for the detection of PCR products in real-time PCR are(1)non-specific fluorescent dyes that intercalate with any double-stranded DNA and (2)sequence-specific DNA probes consisting of oligonucleotides that are labelled with a fluorescent reporter,which permits detection only after hybridization of the probe with its complementary sequence.




Principle

The EMSA/Gel-Shift binding buffer (5X) contains poly(dI-dC) and other active components. The concentration of poly(dI-dC) was optimized to eliminate the non-specific binding between protein and marker probe without weakening the binding between target transcription factor and marker probe.

Applications

For the study of EMSA(gel shift). EMSA can be used to study the binding of target proteins to specific DNA sequences, so that the activation levels of some transcription factors in cells can be studied.

Procedure

1. Sample processing and preparation of PCR reaction system.
2. Add the amplification template, cover the PCR reaction cover, mix well, centrifuge at low speed instantaneously, and transfer to the PCR instrument.
3. Set the program for PCR amplification.
4. Data analysis.

Materials

• Sample Type: serum