Marking method of applying SSAP molecular marker technology to peony

A technology of molecular markers and marker methods, applied in the field of molecular biology, can solve the problems of large heterogeneity and numerous steps of retrotransposon LTR sequences

Inactive Publication Date: 2014-05-14
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

SSAP molecular labeling steps are numerous, and retrotransposon LTR sequences are highly heterogeneous among different species

Method used

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  • Marking method of applying SSAP molecular marker technology to peony
  • Marking method of applying SSAP molecular marker technology to peony
  • Marking method of applying SSAP molecular marker technology to peony

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] A kind of labeling method of SSAP molecular labeling technology on peony, comprises the following steps:

[0105] Step 1. Design primers

[0106] Design Mse I and EcoR I linker primers, pre-amplification PCR primers, linker selective amplification PCR primers, and retroposon PCR primers. The primer sequence direction is from 5' to 3':

[0107] 1) Linker primers

[0108] Mse I Adapter Primer

[0109] Mse I 5′ end adapter primer: Mzx5 GACGATGAGTCCTGAG

[0110] Mse I 3′ end adapter primer: Mzx3 TACTCAGGACTCAT

[0111] EcoR I Adapter Primer

[0112] EcoRI 5′ end adapter primer: Ezx5 CTCGTAGACTGCGTACC

[0113] EcoRI 3′ end adapter primer: Ezx3 AATTGGTACGCAGTCTAC

[0114] 2) Pre-amplification primers

[0115] Mse I preamplification primer: Mzx00 GATGAGTCCTGAGTAAC

[0116] EcoR I preamplification primer: Ezx00 GACTGCGTACCAATTCA

[0117] 3) Selective amplification primers

[0118] Adapter selection amplification primers:

[0119] Mzx02: GATGAGTCCTGAGTAACAT

[0120] Mz...

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Abstract

A marking method of applying an SSAP molecular marker technology to a peony comprises the following steps: designing primers, extracting peony genome DNA, performing enzyme digesting for peony genome DNA, preparing a connector, connecting enzyme digested product with the connector, pre-amplifying of connected product, conducting a selective amplified reaction, testing polyacrylamide gel electrophoresis and the like. According to the marking method, pony genus retrotransposon LTR sequence primer is designed, and through the test of the polyacrylamide gel electrophoresis, stripes are rich; and compared with LTR primer of other species, the primer can directly used to polymorphism analysis between pony genuses, so as to achieve high efficiency and practicability; the selective amplified system is optimized and more suitable for the amplification of pony genus SSAP molecular marker; and silver staining method of polyacrylamide gel electrophoresis is improved, and fixing and dying steps of silver staining are combined to further improve the efficiency of applying the SSAP molecular marker on the pony genus.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a marking method of SSAP molecular marking technology on peony. Background technique [0002] peony( Paeonia suffruticosa Andr.) is a woody plant of the Paeoniaceae Paeoniae group, and is a traditional famous flower native to China. It has rich varieties, huge flowers, colorful and elegant. It is known as "National Beauty and Heavenly Fragrance" and "King of Flowers". It blooms from April to May every year. The flowers are large and colorful, extremely beautiful, elegant and elegant, and the flowers are fragrant, so they are deeply loved by people. Therefore, it is of great significance to improve the research level of peony and promote the industrialization of peony. [0003] Molecular markers are genetic markers based on nucleotide sequence variation in genetic material between individuals, and are a direct reflection of genetic polymorphism at the DNA level. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 侯小改郭大龙张曦宋程威刘素云赵威马慧丽
Owner HENAN UNIV OF SCI & TECH
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