| Autor: |
Karola Marczinek, Peter Nürnberg, Jochen Hampe |
| Rok vydání: |
1999 |
| Předmět: |
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| Zdroj: |
DNA Profiling and DNA Fingerprinting ISBN: 9783764360184 |
| DOI: |
10.1007/978-3-0348-7582-0_13 |
| Popis: |
The basic principle of the two-dimensional (2D) separation of DNA fragments was developed by Fischer and Lehrman in 1979 [1], who applied this method to the detection of variation in the Escherichia coli genome. The introduction of blotting and hybridisation with probes specific for repetitive sequences by Uitterlinden and co-workers made this method applicable to the analysis of complex eukaryotic genomes [2]. 2D DNA fingerprinting is based on separating DNA fragments obtained by digestion with a restriction enzyme according to size and sequence in the first and second dimensions, respectively. Since the melting characteristics of a double-stranded DNA molecule are a function of the base composition of the DNA fragment, this property can be used to achieve a sequence-dependent separation in the second dimension. This is performed in a Polyacrylamide gel containing a linear concentration gradient of the denaturants urea and formamide. Under denaturing conditions, doublestranded DNA fragments migrate through the gel until denaturant concentrations are reached that correspond to the melting temperature of the first domain. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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