Tortoise microsatellite DNA marker based on high-throughput sequencing screening

A technology of DNA labeling and microsatellite labeling, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial measurement/inspection, etc., and can solve problems such as low efficiency and cumbersome work

Active Publication Date: 2017-07-21
ANHUI NORMAL UNIV
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  • Tortoise microsatellite DNA marker based on high-throughput sequencing screening
  • Tortoise microsatellite DNA marker based on high-throughput sequencing screening
  • Tortoise microsatellite DNA marker based on high-throughput sequencing screening

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Embodiment Construction

[0023] The present invention will be described in further detail below in conjunction with the accompanying drawings.

[0024] a) Obtain sequences containing microsatellites

[0025] Turtle genomic DNA was extracted according to the method of sambrook et al. (2002, Molecular Cloning, 3nd463-471), and its integrity was detected by 1% agarose electrophoresis. The DNA sample is sent to the bio company for RAD (Reduced Genome) sequencing.

[0026] The microsatellite sequence was obtained according to the method of Bonatelli et al. (2015, Using Next Generation RAD Sequencing to Isolate Multispecies Microsatellites for Pilosocereus (Cactaceae). 10(11):e0142602).

[0027] b) Design of microsatellite primers

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Abstract

The invention discloses a tortoise microsatellite DNA marker screened based on a high-throughput sequencing method. The high-throughput sequencing method comprises the following steps: establishing a tortoise simplified genomic sequencing library; sequencing; screening microsatellite sequences by using MISA software; and determining 15 abundant-polymorphism microsatellite markers SLN01, SLN02, SLN03, SLN04, SLN05, SLN06, SLN07, SLN08, SLN09, SLN10, SLN11, SLN12, SLN13, SLN14 and SLN15 on the basis of acquisition of a sequence of 987 candidate microsatellites. The invention provides 15 tortoise new microsatellite loca and a primer sequence and method for amplifying the 15 microsatellite loca. The tortoise microsatellite DNA marker can be applied to research of genetic diversity of different populations of tortoises and research of system evolution, and is a reliable and effective molecular marker.

Description

technical field [0001] The invention relates to molecular marker technology, in particular to screening new tortoise microsatellite DNA markers based on high-throughput sequencing. Background technique [0002] In the past few decades, due to the influence of human factors and the change of habitat environment, the population of wild tortoises has declined sharply. At present, this species has been included in the 2004 "China Red Data Book of Endangered Animals" and the 2006 IUCN Red List of Threatened Animal Species. It can be seen that the protection of tortoises is urgent. If we want to formulate effective protection measures for endangered animals, we must first understand the genetic diversity and other information of the species. [0003] Microsatellite (Microsatellite, MS) is also called short tandem repeat sequence or simple repeat sequence. Refers to multiple tandem repeat sequences with a few nucleotides (mostly 2 to 4) as the unit. The repeat unit is more common...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6869C12Q1/6888C12Q2600/156C12Q2535/122C12Q2525/151
Inventor 聂刘旺李军石琼汪玉芹刘谏君王梦
Owner ANHUI NORMAL UNIV
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