Mogurnda adspersa microsatellite markers: multiplexing and multi-tailed primer tagging

Real, Kathryn M.; Schmidt, Daniel J.; Hughes, Jane

doi:10.1007/s12686-009-9095-7

Cited by 35 publications

(39 citation statements)

References 8 publications

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“…Samples were genotyped at six microsatellite loci Cin-1, Cin-10 following (Andreakis et al 2007) and Cin-12, Cin-13, Cin-15, Cin-16 following (Zhan et al 2010). PCRs were performed with the tail method described in (Schuelke 2000), and all forward primers were 5′ modified by adding the Tail 2 sequence from (Real et al 2009). Consequently, the Tail 2 sequence was used as the dye-labeled oligo for each primer pair.…”

Section: Methodsmentioning

confidence: 99%

Oceanographic barriers to gene flow promote genetic subdivision of the tunicate Ciona intestinalis in a North Sea archipelago

et al. 2018

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Pelagic larval development has the potential to connect populations over large geographic distances and prevent genetic structuring. The solitary tunicate Ciona intestinalis has pelagic eggs and a swimming larval stage lasting for maximum a few days, with the potential for a homogenizing gene flow over relatively large areas. In the eastern North Sea, it is found in a geomorphologically complex archipelago with a mix of fjords and open costal habitats. Here, the coastal waters are also stratified with a marked pycnocline driven by salinity and temperature differences between shallow and deep waters. We investigated the genetic structure of C. intestinalis in this area and compared it with oceanographic barriers to dispersal that would potentially reduce connectivity among local populations. Genetic data from 240 individuals, sampled in 2 shallow, and 4 deep-water sites, showed varying degrees of differentiation among samples (FST = 0.0–0.11). We found no evidence for genetic isolation by distance, but two distant deep-water sites from the open coast were genetically very similar indicating a potential for long-distance gene flow. However, samples from different depths from the same areas were clearly differentiated, and fjord samples were different from open-coast sites. A biophysical model estimating multi-generation, stepping-stone larval connectivity, and empirical data on fjord water mass retention time showed the presence of oceanographic barriers that explained the genetic structure observed. We conclude that the local pattern of oceanographic connectivity will impact on the genetic structure of C. intestinalis in this region.Electronic supplementary materialThe online version of this article (10.1007/s00227-018-3388-x) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Oceanographic barriers to gene flow promote genetic subdivision of the tunicate Ciona intestinalis in a North Sea archipelago

et al. 2018

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“…Microsatellite fragments were isolated from genomic DNA following methods in Real et al (2009) with two separate hybridisations of linker-ligated DNA performed using tetramer (dAGAT, dAAGG, dAAAT, dACAT, dAAAG, dAAGT) and dimer/ trimer (dAAC 6 , dAT 15 , dAC 13 , dACC 8 , dAGC 6 , dAAG 8 ) biotinylated oligo probes. Twenty-four fragment inserts per enrichment were sequenced, of which nineteen sequences ranging between 120 and 360 bp were found to contain microsatellite repeats with unique flanking regions.…”

Section: Methodsmentioning

confidence: 99%

“…Genotyping costs were reduced by applying a multi-tailed approach to fluorescent labelling of primers following Real et al (2009). One of four unique 20-mer oligo tails ( Table 2 in Real et al, 2009) was added to the 5′ end of each forward primer, enabling the incorporation of a corresponding fluorescently labelled tagging primer in the PCR (Schuelke, 2000).…”

Section: Methodsmentioning

confidence: 99%

“…One of four unique 20-mer oligo tails ( Table 2 in Real et al, 2009) was added to the 5′ end of each forward primer, enabling the incorporation of a corresponding fluorescently labelled tagging primer in the PCR (Schuelke, 2000). Use of four unique tails permits multiplex amplification with up to four different fluorescent labels in single reaction Marine Genomics 4 (2011) …”

Section: Methodsmentioning

confidence: 99%

“…Marine Genomics j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / m a r g e n (Missiaggia and Grattapaglia, 2006;Real et al, 2009). The four tagging primers were labelled with 6-FAM, VIC, NED and PET from the G5 fluorescent dye set (Applied Biosystems).…”

Section: Contents Lists Available At Sciverse Sciencedirectmentioning

confidence: 99%

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Nine microsatellite markers for the Australian side-necked turtle Chelodina expansa (Chelidae) and cross species amplifications

2011

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Conservation genetics of the Mary River turtle (Elusor macrurus) in natural and captive populations

Schmidt

Espinoza²,

Connell³

et al. 2017

Aquatic Conservation

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1. Many thousands of Mary River turtle eggs were harvested for the pet trade in the 1960s and 1970s before it was recognized as a new species in a unique genus. Pet turtles and their descendants still survive in captive collections. Elusor macrurus is now an endangered species after suffering dramatic population declines along the single Australian river that constitutes its entire range. 2. A conservation genetic assessment was conducted to evaluate population subdivision within the remaining wild population of the Mary River turtle; to compare diversity of the wild population with a captive sample derived from the pet trade; and to establish a baseline estimate of effective population size (N e ) to assist with future monitoring and recovery. 3. Microsatellite analysis indicated panmixia throughout most of the Mary River catchment with the exception of one downstream tributary -Tinana Creek (pop. Specific F ST = 0.154). Subdivision between Tinana Creek and Mary River is a feature common to multiple co-distributed freshwater taxa including the threatened Australian lungfish and Mary River cod. Microsatellite diversity of the wild adult population was low (average H S = 0.554) and not significantly different from that of a sample of captive turtles from the pet tradeindicating genetic diversity may be well represented in captive stocks. Mitochondrial DNA diversity was extremely limited, with only two haplotypes found in the wild and a single shared haplotype in captive turtles.4. Estimates of N e applicable to the entire species in the wild were~136 and~158 using two independent methods. A reasonable management objective should be retention of N e levels >100 during recovery of the species. Additional recommendations include that Mary River turtles be listed as Critically Endangered, and that a recovery plan be developed that considers 'headstarting'using captive bred stocks to supplement the wild population.

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Mogurnda adspersa microsatellite markers: multiplexing and multi-tailed primer tagging

Cited by 35 publications

References 8 publications

Oceanographic barriers to gene flow promote genetic subdivision of the tunicate Ciona intestinalis in a North Sea archipelago

Oceanographic barriers to gene flow promote genetic subdivision of the tunicate Ciona intestinalis in a North Sea archipelago

Nine microsatellite markers for the Australian side-necked turtle Chelodina expansa (Chelidae) and cross species amplifications

Conservation genetics of the Mary River turtle (Elusor macrurus) in natural and captive populations

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