Primed Synthesis and Direct Sequencing in the Isolation of cDNA Clones Using Short Oligonucleotides
With the development of improved techniques for the construction of cDNA librari
With the development of improved techniques for the construction of cDNA libraries representing rare messenger species has come increased demand for screening techniques to isolate specific cDNAs. A variety of techniques has been developed:
| 1. | The use of exact and degenerate oligonucleotides complementary to RNAs coding for established protein sequence. |
| 2. | Bacterial expression of cDNA and immunological detection of the protein products. |
| 3. | Selection by hybrid-arrest or hybrid-select translation. |
| 4. | Differential hybridization using cDNAs obtained under different physiological conditions. |
| 5. | Bacterial expression of cDNA and selection by biological assay. |
| 6. | Transfer of cDNA libraries in eukaryotic expression vectors into mammalian cell lines and subsequent biological selection. |
| 7. | Expression of cloned cDNAs and mRNA in SP6 vectors with subsequent microinjection of RNA and biological selection of decreasing pools. |
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