A hands-on overview of tissue preservation methods for molecular genetic analyses

Nagy, Zoltán T.

doi:10.1007/s13127-010-0012-4

Cited by 121 publications

(150 citation statements)

References 90 publications

Supporting

Mentioning

135

Contrasting

Unclassified

Order By: Relevance

“…Although we cannot prove this statistically, the cooling chain and fresh tissue may be regarded as essential for a high success rate. However, there is evidence (not shown here) suggesting that ac− ceptable results might be possible even after two years of storage as long as the samples were constantly kept in chilled conditions as recommended for various other taxa (Quicke et al 1999;Gemeinholzer et al 2010;Nagy 2010).…”

Section: Discussionmentioning

confidence: 75%

Field and Laboratory Methods for DNA Studies on Deep-sea Isopod Crustaceans

Riehl¹,

Brenke²,

Brix³

et al. 2014

Polish Polar Research

View full text Add to dashboard Cite

Field and laboratory protocols that originally led to the success of published stud− ies have previously been only briefly laid out in the methods sections of scientific publica− tions. For the sake of repeatability, we regard the details of the methodology that allowed broad−range DNA studies on deep−sea isopods too valuable to be neglected. Here, a com− prehensive summary of protocols for the retrieval of the samples, fixation on board research vessels, PCR amplification and cycle sequencing of altogether six loci (three mitochondrial and three nuclear) is provided. These were adapted from previous protocols and developed especially for asellote Isopoda from deep−sea samples but have been successfully used in some other peracarids as well. In total, about 2300 specimens of isopods, 100 amphipods and 300 tanaids were sequenced mainly for COI and 16S and partly for the other markers. Although we did not set up an experimental design, we were able to analyze amplification and sequencing success of different methods on 16S and compare success rates for COI and 16S. The primer pair 16S SF/SR was generally reliable and led to better results than univer− sal primers in all studied Janiroidea, except Munnopsidae and Dendrotionidae. The widely applied universal primers for the barcoding region of COI are problematic to use in deep−sea isopods with a success rate of 45-79% varying with family. To improve this, we recommend the development of taxon−specific primers.

show abstract

Section: Discussionmentioning

confidence: 75%

Field and Laboratory Methods for DNA Studies on Deep-sea Isopod Crustaceans

Riehl¹,

Brenke²,

Brix³

et al. 2014

Polish Polar Research

View full text Add to dashboard Cite

show abstract

“…Although samples collected with FTA card were immediately stabilised within the card's cellulose matrix they still required an additional step of drying before storage for transport. However, once dried, FTA cards can provide a validated DNA archive that could be stored at room temperature for an extended period of time (Smith and Burgyone 2004;Nagy 2010;Anon. 2011).…”

Section: Discussionmentioning

confidence: 99%

“…These reagents increase the individual unit sampling costs, but reduce the shipping logistics. In addition, the long-term security of the samples to a similar level as provided by FTA cards can be accomplished if the samples are frozen (Nagy 2010). Alternative solutions are under investigation to further reduce the cost of stabilisation of the tissue in the GTT tip.…”

Section: Discussionmentioning

confidence: 99%

A new tool in the toolbox for large-scale, high-throughput fisheries mark-recapture studies using genetic identification

Bradford

Hill

Davies

et al. 2016

Mar. Freshwater Res.

View full text Add to dashboard Cite

Abstract. The lack of independently verifiable estimates of catches and fisheries independent estimates of abundance and fishing mortality are major sources of uncertainty in the management of many fisheries. DNA profiling provides the potential to substantially improve the quality of data for assessments and act as an additional deterrent to illegal, unreported, and unregulated (IUU) fishing. Barriers to the implementation of this technology include cost of sample collection and processing, forensic grade quality control, and the ability to apply undetectable tags. We present the results of a comparison of two current and one new (gene tag tool, GTT) sampling techniques, using the highly valued southern bluefin tuna as an example. We demonstrate that fish sampled with two techniques are highly unlikely to be recognised as 'tagged', whereas one technique was easily recognisable after 73 days. The GTT reduced handling before DNA extraction, whereas both other techniques require additional labour, adding to cost and potential contamination of the evidentiary chain. Evidence of cross-contamination in the Whatman FTA Elute samples suggests they may not be as suitable for at-sea field applications. Two of the three sampling techniques are capable of obtaining high quality tissue samples for stock assessment and chain of custody purposes in a cost-effective and unidentifiable manner.

show abstract

“…specimen age, dehydration technique) for DNA recovery (Junqueira et al 2002). Nevertheless, the speed at which the internal tissues dry can affect the ultimate state of preservation of DNA for subsequent genomic extraction (Quicke et al 1999;Nagy 2010). The HMDS dehydration method, along with other chemical-based techniques, is demonstrated to yield high quality genomic DNA (Austin and Dillon 1997); some DNA sequence data in Buffington et al (2007), using the chelex extraction protocol, was generated from cynipoid specimens that were vacuum dried in the manner summarized here, but no quantification of success vs. failure of DNA amplification was documented.…”

Section: Discussionmentioning

confidence: 99%

Description of two techniques to increase efficiency in processing and curating minute arthropods, with special reference to parasitic Hymenoptera

Gates¹,

Buffington²

2011

JHR

View full text Add to dashboard Cite

We describe and illustrate two techniques for enhancing curatorial and processing efficiency as it pertains to parasitic Hymenoptera (Chalcidoidea, Cynipoidea). These techniques were developed in response not only to the massive number of parasitoids that have been acquired through various biodiversity studies, but also the difficulty in mobilizing the human resources to curate this material. The first technique uses small, crystal polystyrene boxes with tight-fitting lids to store dehydrated specimens prior to mounting. Locality information is affixed to the box and specimens are spread in a layer for ease of examination by researchers. Solutions for managing static electricity within the specimen boxes are discussed. The second involves a vacuum pump connected to a funnel with a filtration membrane and flask apparatus to rapidly dehydrate hard-bodied parasitoids (Figitidae) that are not subject to collapse during air-drying.

show abstract

A hands-on overview of tissue preservation methods for molecular genetic analyses

Cited by 121 publications

References 90 publications

Field and Laboratory Methods for DNA Studies on Deep-sea Isopod Crustaceans

Field and Laboratory Methods for DNA Studies on Deep-sea Isopod Crustaceans

A new tool in the toolbox for large-scale, high-throughput fisheries mark-recapture studies using genetic identification

Description of two techniques to increase efficiency in processing and curating minute arthropods, with special reference to parasitic Hymenoptera

Contact Info

Product

Resources

About