Isolation of Genomic Insertion Sites of Proviruses Using Splinkerette-PCR-Based Procedures
The availability of whole genomic sequences provides a great framework for biologists to address a broad range of scientific questions. However, functions of most mammalian genes remain obscure. The forward genetics strategy of insertional mutagenesis uses DNA mutagens such as retroviruses and transposable elements; this strategy represents a powerful approach to functional genomics. A variety of methods to uncover insertion sites have been described. This chapter details SplinkTA-PCR and SplinkBlunt-PCR, modified from splinkerette-PCR, for mapping chromosomally the insertion sites of a murine leukemia virus that causes leukemia in the BXH-2 strain of mice. These protocols are easy to use, reliable, and efficient.
- 差示反转录PCR[mRNA差异显示技术]
- Antibody Development and Use in Chromogenic and Fluorescent Immunostaining
- Challenges of Sticky Co-immunoprecipitation: Polyalanine Tract ProteinProtein Interactions
- In Vitro Translation Using Rabbit Reticulocyte Lysate
- Detecting MicroRNA in Human Cancer Tissues with Fluorescence In Situ Hybridization
- A High-throughput Gateway-Compatible Yeast One-Hybrid Screen to Detect ProteinDNA Interactions
- The Use of Artificial MicroRNA Technology to Control Gene Expression in Arabidopsis thaliana
- Application of Microarrays for DNA Methylation Profiling
- In Silico PCR Analysis
- Epitope Mapping Using Homolog-Scanning Mutagenesis