PCR Method for Generating Multiple Mutations at Adjacent Sites
Site-directed mutagenesis is a commonly used tool for identifying the role of specific amino acids in the structure and function of proteins. Various methods of in vitro mutagenesis have been described and are widely used for introducing modified coding sequences (1 -7 ). In comparison, polymerase chain reaction (PCR)-based methods (1 -6 ) are generally faster and more efficient than non-PCR-based methods (7 ). On the other hand, some PCR-based methods require two or more primers for each round of mutagenesis, whereas others need a single very long oligonucleotide or two round of PCR for introducing one mutation (5 -7 ). These all can increase the cost of mutagenesis.
- Methylation-Specific PCR In Situ Hybridization
- Mitochondrial D-Loop and Coding Sequence Analysis Using Pyrosequencing
- Design Considerations for Genetic Linkage and Association Studies
- An RT-PCR-Based Protocol for the Rapid Generation of Large, Representative cDNA Libraries for Expression Screening
- RNA-Seq Analysis of Gene Expression and Alternative Splicing by Double-Random Priming Strategy
- In Vitro Reconstitution of In Vivo-Like Nucleosome Positioning on Yeast DNA
- Clinical Production and Therapeutic Applications of Alloreactive Natural Killer Cells
- Agrobacterium-Mediated Transformation of Bread and Durum Wheat Using Freshly Isolated Immature Embryos
- Fluorescent Detection of Telomerase Activity
- Optimization of Purification Protocols Based on the Step-by-Step Monitoring of the Protein Aggregates in Soluble Fractions