A set of mini-barcoding primers for obtaining DNA barcodes of beetle specimens in collections and its application

Abstract

The invention discloses a set of Mini-Barcoding primers for obtaining DNA barcode sequences of beetle specimens in collections, consisting of primer pairs Cocc301, Cocc286 and Cocc214 or primer pairs Cocc301 and Cocc413 or primer pairs Cocc301, Cocc286, Cocc214 and Cocc658 or primer pairs Cocc301, Cocc413 and Cocc658. Using ordinary PCR or nested PCR technology, two or three target gene fragments of beetle specimens are amplified, and then spliced ​​into a complete collection of beetle specimen DNA barcode sequences. The mini barcode primer designed by the invention has good amplification efficiency, overcomes the problem of low amplification efficiency of traditional barcode primers, and has extremely high amplification efficiency especially for long-lived library specimens. The primer designed by the invention can Efficiently obtain the DNA barcode sequences of beetle specimens in the collection, enrich the beetle DNA barcode database, and provide a reference for the identification of beetles using DNA barcodes.

Description

technical field [0001] The invention belongs to the technical field of insect molecular identification, and more particularly, relates to a set of Mini-Barcoding primers for obtaining DNA barcodes of beetle specimens in museum collections and applications thereof. Background technique [0002] Beetles are the collective name for insects of the order Coleoptera. At present, there are more than 380,000 species of beetles in the world, accounting for 40% of the total number of insects, and it is the largest group of insects. The rich biodiversity of beetles enables them to play an important role in terrestrial and freshwater ecosystems. To adapt to diverse habitats, beetles exhibit extreme diversity in body size, shape, and structure. Therefore, it is very difficult to identify and distinguish beetles by traditional morphological characteristics, especially in some taxa for which traditional taxonomy is not perfect. Therefore, it is particularly important to identify and dif...

AI Technical Summary

Problems solved by technology

[0004] Because collecting fresh samples is not only time-consuming and laborious, but also difficult to cover a comprehensive range of species, when obtaining the DNA barcode sequence of ladybugs, collection specimens are often used

Since the usual method of preserving insect specimens is needle insertion, the DNA in the specimen will degrade and fragment over time, so it is difficult to use universal primers to obtain the complete DNA barcode sequence

In order to solve this problem, Van Houdt et al. (2010) proposed to design new universal primers to amplify several short fragments with a length of 269-363bp, and then connect them together to obtain the full-length sequence of the barcode. The problem of low efficiency, and it cannot be used to detect specimens that have been stored for a long time

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