The probeless detection of hidden genetic variation by Restriction site generating-polymerase chain reaction (RG-PCR) (CAT#: STEM-MT-0011-LGZ)

Introduction

RG-PCR was developed and applied to screen for several relatively common cystic fibrosis mutations that cannot be analyzed with known restriction enzymes: G542X, 2869insG, Y913C, N1303K, and 1717-1GA. This method can identify almost any single base change by restriction endonuclease analysis, does not require molecular probes, is fast and easy to perform.

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Principle Applications Procedure Materials Notes Related Products

Principle

We have a DNA amplification technique in which modified primers introduce a base substitution near the codon of interest and create an artificial restriction site for detection of mutations that do not create or modify naturally occurring restriction sites (restriction site generating-polymerase chain reaction, RG-PCR).

Applications

Application to the study of some common cystic fibrosis mutations

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: depends on the customer's analysis requirements

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