Introduction

Generation Mice

The genomes of humans and mice have been completely sequenced and about 30,000 genes have been detected in both organisms 1-3 . The challenge is now to elucidate the function of all these genes. The most powerful way to perform such functional genomics is gene targeting, allowing the targeted overexpression, deletion, or mutation of a gene product in a mouse and thereby studying its function in the context of all other unaffected genes. During the 15 yr in which this technology has been...

Basic Principles and Routine Practice Yanbin Dong and Haidong

Mobility Shift Sscp

Single-strand conformational polymorphism SSCP analysis is a simple and sensitive technique for mutation detection and genotyping. The principle of SSCP analysis is based on the fact that single-stranded DNA has a defined conformation. Altered conformation due to a single base change in the sequence can cause single-stranded DNA to migrate differently under nondenaturing electrophoresis conditions. Therefore wild-type and mutant DNA samples display different band patterns. SSCP analysis...

Denaturing High Performance Liquid Chromatography Using the Wave Dna Fragment Analysis System

Heteroduplex Dna

Denaturing high-performance liquid chromatography DHPLC is a chromatographic mutation analysis technique that is based on temperature-dependent separation of DNA containing mismatched base pairs from polymerase chain reaction PCR -amplified DNA fragments. The WAVE system, developed for DHPLC analysis, allows for unattended analysis of 96 samples directly from a PCR plate under a number of different conditions. It utilizes a Peltier cooling platform to maintain sample integrity. Sample detection...