1995 GfE Goettingen: DNA fingerprinting in C elegans and related species
Source: Tagungsband der 11. Wissenschaftlichen Tagung der Gesellschaft für Entwicklungsbiologie e. V. vom 21. - 23. März 1995 in Göttingen (GER)
[More articles from MB] [Articles about MB]
FROM MARK BENECKE and EINHARD SCHIERENBERG
We are interested in phylogenetic relationships among nematodes and
related taxa. Conventional multiloeus fingerprinting and the PCR-based RAPD
technology of DNA fingerprinting appear to be promising tools for a molecular phylogenetic
approach: They allow strain-up to genus-specific characterization of nematodes.
a. Multilocus fingerprinting Using short synthetic oligonucleotides, we hybridized
against Southern blots of digested genomic DNA. The oligonucleotide probes we
selected detect a set of tandem repeated non-coding sequences of variable
length.
b. PCR fingerprinting Genomic DNA from the C. elegans wildtype and a C. elegans
isolate from Australia, from another nematode genus (Rhabditis) and from a turbellarian
species were subjected to PCR using several short synthetic oligonucleotides. Only a single oligonucleotide primer is employed per PCR reaction.
These short primers, like short oligonucleotides in multiloeus fingerprinting,
visualize DNA sites which are located close to one another in inverted orientation.
The technique essentially scans a genome for these small inverted repeats
and amplifies intervening DNA segments of variable length (Random Amplified
Polymorphie DNA).
This technique requires an extremely low amount of DNA
(DNA of a single C. elegans individual appears to be sufficient for several RAPD
experiments).
Depending on the degree of variability, probes and primers should be useful
for studies on different taxonomie levels. One advantage of DNA fingerprinting over
sequencing of specific RNA's or genes is that information spread over the entire
genome is obtaines in one working step.
Our fingerprints indicate that individual strains can be distinguished by their
defined banding patterns. In conjunction with developmental studies we plan to use
this assay to analyze phylogenetic relationships among nematodes. In addition, we
would like to explore to which extent the RAPD technology is suitable for comparative
studies on higher taxonomic levels.
suggested readings
- More about DNA and DNA typing ENGLISH AND GERMAN TEXT
- Katzenhaar klärt Mord auf GERMAN TEXT
- Urinproben auf der Spur GERMAN TEXT
- Heulen hilft nicht GERMAN TEXT
- Der genetische Fingerabdruck (skilled work) GERMAN TEXT