2017
DOI: 10.1002/ece3.3212
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New specific primers for amplification of the Internal Transcribed Spacer region in Clitellata (Annelida)

Abstract: Nuclear molecular evidence, for example, the rapidly evolving Internal Transcribed Spacer region (ITS), integrated with maternally inherited (mitochondrial) COI barcodes, has provided new insights into the diversity of clitellate annelids. PCR amplification and sequencing of ITS, however, are often hampered by poor specificity of primers used. Therefore, new clitellate‐specific primers for amplifying the whole ITS region (ITS: 29F/1084R) and a part of it (ITS2: 606F/1082R) were developed on the basis of a coll… Show more

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Cited by 8 publications
(4 citation statements)
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“…Genomic DNA was extracted from the dried fruiting body of mushrooms using the cetyl trimethyl ammonium bromide (CTAB) method [ 25 ]. Highly conserved regions of fungal rDNA-specific genes (i.e., ITS1 and ITS4) were amplified using the PCR amplification method [ 26 ]. During PCR amplification, ITS1-F (5′-CTT GGT CAT TTA GAG GAA GTA A-3′) and ITS-4 (5′-TCC TCC GCT TAT TGA TAT GC-3′) set of forward and reverse primers were used, respectively ( Table 2 ).…”
Section: Methodsmentioning
confidence: 99%
“…Genomic DNA was extracted from the dried fruiting body of mushrooms using the cetyl trimethyl ammonium bromide (CTAB) method [ 25 ]. Highly conserved regions of fungal rDNA-specific genes (i.e., ITS1 and ITS4) were amplified using the PCR amplification method [ 26 ]. During PCR amplification, ITS1-F (5′-CTT GGT CAT TTA GAG GAA GTA A-3′) and ITS-4 (5′-TCC TCC GCT TAT TGA TAT GC-3′) set of forward and reverse primers were used, respectively ( Table 2 ).…”
Section: Methodsmentioning
confidence: 99%
“…Two pairs of primers were tested: the commonly used pair mrSSU1 and mrSSU3R, and our newly designed mycobiont-specific primer pair mt-SSU-581-5’ and mt-SSU-1345-3’. In the setting of simulations, amplicon sizes were accepted between 200 bp to 2500 bp , and only up to three nucleotide mismatches between primers and templates were allowed (except for the last two positions at the 3’ end), according to described parameters ( Bellemain et al 2010 ; Riaz et al 2011 ; Liu and Erséus 2017 ).…”
Section: Methodsmentioning
confidence: 99%
“…Internal transcribed spacer (ITS) gene sequences of Artemisia were collected for phylogenetic analysis. The ITS serves as a research region in phylogeny, biogeography, and population genetics ( Liu and Christer, 2017 ; Ren et al, 2017 ). Chrysanthemum , Atractylodes , and Cirsium in Asteraceae were selected as outgroups as they are relatively common genera in Asteraceae and are commonly used as medicinal plants.…”
Section: Methodsmentioning
confidence: 99%