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Primer combination for constructing wolfberry DNA fingerprint spectrum, and applications and method

A technology of DNA fingerprinting and primer combination, which is applied in the field of DNA fingerprinting, can solve the problems that DNA fingerprinting has not been reported, and the application research of DNA fingerprinting is not much, so as to achieve the effect of accurate and reliable fingerprint information

Active Publication Date: 2017-05-31
WOLFBERRY ENG RES INST NINGXIA ACADEMY OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are not many studies on the application of DNA fingerprints on Lycium barbarum, especially the use of SSR molecular marker technology to construct Lycium barbarum DNA fingerprints has not been reported.

Method used

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  • Primer combination for constructing wolfberry DNA fingerprint spectrum, and applications and method
  • Primer combination for constructing wolfberry DNA fingerprint spectrum, and applications and method
  • Primer combination for constructing wolfberry DNA fingerprint spectrum, and applications and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090]The primer combination provided in this example for constructing Lycium barbarum DNA fingerprints includes primer pairs 1-15, and the base sequences of the upstream primer (F) and downstream primer (R) of each primer pair are shown in Table 1 below. Wherein, the 5' end of the upstream primer of each primer pair is connected with an M13 universal primer, and its sequence is shown in the underlined part in Table 1. Of course, in other embodiments, the M13 universal primer may not be connected.

[0091] Table 1. The base sequence of the 1-15th primer pair used to construct Lycium barbarum DNA fingerprints provided in this example

[0092]

[0093]

[0094] The primers named SF1, SF3, SF5, SF12, SF13, SF14, SF15, SF20, SF30, SF51, SF61, SF63, SF68, SF92, and SF107 in Table 1 correspond to the first primer pair, the second primer pair, and the third primer pair respectively. Primer pair, 4th primer pair, 5th primer pair, 6th primer pair, 7th primer pair, 8th primer pai...

Embodiment 2

[0110] Due to the common agarose gel electrophoresis used in Example 1 to achieve the effect of separating PCR amplification products, it has low resolution, and the separation effect for bands with different base numbers is not obvious, which easily leads to the obtained The phenomenon that the DNA fingerprint results expressed by electrophoresis are inaccurate. Therefore, for the purpose of further perfecting and improving the invention, this embodiment carries out fluorescent gene labeling on the primer combination used to construct the Lycium barbarum DNA fingerprint, so that the primer combination used to construct the Lycium barbarum DNA fingerprint provided in this example, The amplified product can be used for capillary gel electrophoresis to improve the resolution of the electrophoresis spectrum and ensure that the obtained DNA fingerprint spectrum is more accurate.

[0111] The combination of primers provided in this example for constructing the Lycium barbarum DNA f...

Embodiment 3

[0114] The combination of primers provided in this example for constructing the Lycium barbarum DNA fingerprint includes primer pairs 1-15. The 5' end of the upstream primer of each primer pair in the 1-15 primer pair is marked with a fluorescent gene, wherein the fluorescent gene of the upstream primer of the 1-10 primer pair and the 13th primer pair is TAMRA; the 11th primer pair, The fluorescent gene of the upstream primers of the 12th primer pair, the 14th primer pair, and the 15th primer pair is HEX.

[0115] The base sequences of the upstream primer (F) and the downstream primer (R) of each primer pair in the 1-15th primer pair provided in this example are the same as those in Example 1.

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PUM

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Abstract

The invention provides a primer combination for constructing a wolfberry DNA fingerprint spectrum, and applications and a method, belonging to the technical field of a DNA fingerprint spectrum. The method comprises the following steps: on the basis of wolfberry gene sequencing results, firstly developing and screening out 15 SSR primer pairs with polymorphism, wherein the 15 SSR primers can be used for constructing the wolfberry DNA fingerprint spectrum; and constructing the wolfberry DNA fingerprint spectrum by combining the capillary electrophoresis technology. The method has the characteristics of being high in electrophoresis resolution and sensitivity, portable in data analysis and processing and the like, thus providing a solid theory support for further application research of the DNA fingerprint spectrum on wolfberries.

Description

technical field [0001] The invention relates to the technical field of DNA fingerprints, in particular to a combination of primers for constructing Lycium barbarum DNA fingerprints as well as an application and a method. Background technique [0002] Lycium barbarum is a traditional Chinese herbal medicine in my country and plays an important role in the field of Chinese herbal medicines. As an authentic wolfberry producing area, Ningxia has a long history of wolfberry cultivation and has achieved fruitful results in wolfberry breeding, cultivation, and processing. In recent years, with the continuous improvement of wolfberry breeding methods, new wolfberry varieties have also emerged, and many excellent varieties of these wolfberry germplasm resources have not been rationally utilized. Therefore, effective classification and identification of these wolfberry germplasm resources is particularly important for the research and variety innovation of wolfberry germplasm resourc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 曹有龙尹跃赵建华安巍
Owner WOLFBERRY ENG RES INST NINGXIA ACADEMY OF AGRI & FORESTRY SCI
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