|Table of Contents|

The Optimization of SRAP-PCR Amplification Program and System in Kale(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2014年09期
Page:
121-124
Research Field:
Publishing date:

Info

Title:
The Optimization of SRAP-PCR Amplification Program and System in Kale
Author(s):
ZHU Peng-fang1; KANG Yao-hai1; HUANG Juan-juan1; FANG Xia1; LIU Chang1; ZHAO Ying1; 2
1.College of Forestry,Shenyang Agricultural University,Shenyang,Liaoning 110866；
2.Liaoning Urban Construction Technical College,Shenyang,Liaoning 110122
Keywords:
kale; SRAP marker; system optimization
PACS:
S 635
DOI:
-
Abstract:
Taking the genomic DNA extracted from kale leaves as template,each influencing factor of amplified polymorphism polymerase chain reaction (SRAP-PCR) related to the sequence was set by the single factor gradient and the program was optimized,the SRAP which were high polymorphism,good repeatability,clear were screened.The results showed that the optimized protocol was as follows:a total volume of 10 μL containing 50 ng genomic DNA,1×buffer,0.20 mmol/L dNTPs,1 U of Taq DNA polymerase and 0.60 μmol/L primers.Protocol run under the following conditions:predenaturing at 94℃ for 5 min,then denatured at 94℃ for 30 s,annealed at 35℃ for 30 s,and an extension at 72℃ for 30 s for 5 times,denatured at 94℃ for 30 s and annealed at 50℃ for 30 s,and an extension at 72℃ for 30 s for 35 times,extension at 72℃ for 7 min at last,then kept at 4℃.The above optimal SRAP-PCR reaction system and amplification procedure were checked by 22 of F2 individuals.The system and procedure could be used in genomic DNA SRAP-PCR amplification in kale.

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Last Update: 2014-08-13