|Table of Contents|

Optimization of AFLP Reaction System and Screening of Primers for Pogostemon cablin

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2020年11
Page:
117-123
Research Field:
Publishing date:

Info

Title:
Optimization of AFLP Reaction System and Screening of Primers for Pogostemon cablin
Author(s):
LIU Xiaoying1; LU Changhua1; LI Jiahui1; YAN Hanjing1; 2; 3; ZHANG Hongyi1; 2; 3; HE Mengling1; 2; 3
(1.School of Traditional Chinese Medicine,Guangdong Pharmaceutical University，Guangzhou,Guangdong 510006；2.Key Laboratory of State Administration of Traditional Chinese Medicine for Production & Development of Cantonese Medicinal Materials,Guangzhou,Guangdong 510006；3.Comprehensive Experimental Station of Guangzhou,Chinese Materia Medica,China Agriculture Research System,Guangzhou,Guangdong 510006)
Keywords:
Pogostemon cablin; AFLP molecular marker; system optimization; primer screening
PACS:
-
DOI:
10.11937/bfyy.20193400
Abstract:
The Pogostemon cablin leaves were used as the test material and the AFLP molecular labeling method was used to study the AFLP reaction system of Pogostemon cablin,including key factors such as reaction system of digestion and ligation dilution multiple of ligation product and pre-amplification product,and then screened the AFLP primer combinations (E/M combinations) in order to lay the foundation for the subsequent screening of superior germplasm resources of P.cablin.The results showed that the digestion system was 20 μL,containing 300 ng of template DNA,and reacted at 37 ℃ for 3 hours；the ligation reaction was carried out at 16 ℃ for 12 hours；the optimal dilution multiple of the ligation product was 3 times；the optimal dilution multiple of the pre-amplified product was 20 times.12 pairs of primer combinations with clear and polymorphic bands were screened by optimized reaction system.The optimized reaction system was suitable for the AFLP analysis of P.cablin,and the selected primers also had good polymorphism,which can provide technical support for the subsequent screening of excellent germplasm of P.cablin.

References:

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Last Update: 2020-08-12