Nematode‐specific PCR primers for the 18S small subunit rRNA gene

Abstract

A set of polymerase chain reaction primers were designed, which amplify a c. 1 kb fragment of the 18S ribosomal DNA gene, and are specific to the phylum Nematoda. These have proven useful in isolating nematode genes from samples mixed with other biological material, particularly with application to DNA barcoding. Optimal reaction conditions are described. These primers have successfully amplified the correct fragment from a wide phylogenetic range of nematodes, and have so far generated no sequences from any other organismal group.

Morphological and molecular characterization of Cucullanus hainanensis sp. nov. (Ascaridida: Cucullanidae) from Muraenichthys gymnopterus (Bleeker) (Anguilliformes: Ophichthidae) in the South China Sea

“…DNA was eluted in elution buffer and kept at -20°C until use. The partial 18S rDNA was amplified by PCR using the primers 18SF (forward: 5'-CGCGAATRGCTCATTACAACAGC-3') and 18SR (reverse: 5'-GGGCGGTATCTGATCGCC-3') (Floyd et al 2005). The ITS-2 region was amplified by PCR using the primers XZ1 (forward: 5'-ATTGCGCCATCGGGTTC ATTCC-3') and NC2 (reverse: 5'-TTAGTTTCTTTTCCTC-CGCT-3') (Zhu et al 2002).…”

Wolbachia Are Present in Southern African Scorpions and Cluster with Supergroup F

“…Sequences were deposited at the MLST website. All samples were screened using nematode-specific 18S primers [10] to exclude contamination with parasitic nematode DNA (and potential associated Wolbachia symbionts). Results were negative.…”

New molecular data for parasites Hammerschmidtiella indicus and Thelandros scleratus (Nematoda: Oxyurida) to infer phylogenetic position

“…Extraction of genomic DNA for both species was performed on individual female nematodes using the DNeasy Tissue Kit from QIAGEN according to the manufacturer's protocol. The primer set Nem 18S F (5'-CGCGAATRGCTCATTACAACAGC-3') and Nem 18S R (5'-GGGCGGTATCTGATCGCC-3') designed by Floyd et al (2005) was used for amplification of the region of 18S ribosomal DNA. PCR was performed in a total volume of 25 µL, comprising 1X PCR buffer with MgCl 2 , a mixture of dNTPs (at a concentration of 0.4 mM), primers at 0.8 µL each, 1 U Taq DNA polymerase (Biotools, Madrid, Spain), and 3 µL of extracted nematode DNA template.…”