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Normalization of Full-Length-Enriched cDNA
A well-recognized obstacle to efficient high-throughput analysis of cDNA libraries is the differential abundance of various transcripts in any particular cell type. Decreasing the prevalence of clones representing abundant transcripts before sequencing, using cDNA normalization, may significantly increase the efficacy of random sequencing and is essential for rare gene discovery. Duplex-specific nuclease (DSN) normalization allows the generation of normalized full-length-enriched cDNA libraries to permit a high gene discovery rate. The method is based on the unique properties of DSN from the Kamchatka crab and involves denaturation–reassociation of cDNA, degradation of the ds-fraction formed by abundant transcripts by DSN, and PCR amplification of the remaining ss-DNA fraction. The method has been evaluated in various plant and animal models.
- Comparative Molecular Physiological Genomics: Heterologous Probing of cDNA Arrays
- MethylQuant: A Real-Time PCR-Based Method to Quantify DNA Methylation at Single Specific Cytosines
- Solid Phase Fluorescent Sequencing of the CFTR Gene
- Melting Curve Assays for DNA Methylation Analysis
- PCR Diagnosis of the Bovine Immunodeficiency-Like Virus
- Techniques for the Isolation and Use of Conditionally Expressed Antisense RNA to Achieve Essential Gene Knockdowns in Staphyloco
- Methylation Analysis by Microarray
- Overlap Extension PCR: An Efficient Method for Transgene Construction
- Cloning of ES Cells and Mice by Nuclear Transfer
- Clustering of Gene Expression Data Via Normal Mixture Models
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