PCR based targeted genomic and cDNA differential display
| Autor: | CassandraL. Smith, Sergey Lukyanov, JoelH. Graber, NataliaE. Broude, Eugene D. Sverdlov, Sarah Malpel, Niels Storm |
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| Rok vydání: | 1999 |
| Předmět: |
Genetics
Differential display DNA Complementary Interspersed repeat Biology Applied Microbiology and Biotechnology Genome Polymerase Chain Reaction law.invention Conserved sequence genomic DNA chemistry.chemical_compound Huntington Disease chemistry Trinucleotide Repeats law Complementary DNA Humans Polymerase chain reaction DNA Alleles |
| Zdroj: | Genetic analysis : biomolecular engineering. 15(2) |
| Popis: | We previously described a targeted genomic differential display method (TGDD: Broude NE, Chandra A, Smith CL. Differential display of genomic subsets containing specific interspersed repeats. Proc. Natl. Acad. Sci. USA 1997;94:4548–53). In that method, presently characterized as method I, targeting was accomplished by capturing DNA fragments containing specific a sequence by hybridization with complementary single-stranded DNA. The captured fragments were amplified by PCR. Here, we describe method II where targeting is accomplished by PCR using primers specific to the target sequence. Method II takes advantage of PCR suppression to eliminate fragments not containing the target sequence (Siebert PDA, Chenchik A, Kellogg DE, Lukyanov KA and Lukyanov SA. An improved PCR method for walking in uncloned genomic DNA. Nucleic Acids Res 1995;23:1087–1088). Targeting focuses analysis on and around interesting areas and additionally serves to reduce the complexity of the amplified subset. These approaches are useful to amplify genome subsets containing a variety of targets including various conserved sequences coding for cis -acting elements or protein motifs. |
| Databáze: | OpenAIRE |
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