Volume 27 Issue 6
Dec.  2010
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WANG Hong-ling, ZHANG Jue, JIAO Zhong-yi, SHI De-zong. Establishing and optimizing the SRAP-PCR system for Salix matsudana[J]. Journal of Zhejiang A&F University, 2010, 27(6): 945-949. doi: 10.11833/j.issn.2095-0756.2010.06.024
Citation: WANG Hong-ling, ZHANG Jue, JIAO Zhong-yi, SHI De-zong. Establishing and optimizing the SRAP-PCR system for Salix matsudana[J]. Journal of Zhejiang A&F University, 2010, 27(6): 945-949. doi: 10.11833/j.issn.2095-0756.2010.06.024

Establishing and optimizing the SRAP-PCR system for Salix matsudana

doi: 10.11833/j.issn.2095-0756.2010.06.024
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  • Corresponding author: WANG Hong-ling
  • Publish Date: 2010-12-20
通讯作者: 陈斌, bchen63@163.com
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    沈阳化工大学材料科学与工程学院 沈阳 110142

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Establishing and optimizing the SRAP-PCR system for Salix matsudana

doi: 10.11833/j.issn.2095-0756.2010.06.024
    Corresponding author: WANG Hong-ling

Abstract: To enrich the molecular marker method for willow trees,an improved reaction system from a sequence-related amplified polymorphism-polymerase chain reaction (SRAP-PCR) of Salix matsudana(Hankow willow) was obtained with an orthogonal design. In addition,the temperatures for two annealing methods were optimized in the PCR amplifier and a polyacrylamide gel electrophoresis(PAGE) was employed. Results showed that the reaction system was 20.84 nkat Taq,2.00 L Mg2+(25.00 mmolL-1),2.50 L deoxyribonucleotide triphosphates (dNTPs)(2.50 mmolL-1 each),1.50 L primer(20.00 molL-1),100.00 ng template,and 2.50 L 10 Taq buffer in 25.00 L. The program was as follows:94 ℃ for 2.0 min;then,for 5 cycles,94 ℃ for 1.0 min,38 ℃ for 1.0 min,and 72 ℃ for 1.5 min;next,at 30 cycles,94 ℃ for 1.0 min,54 ℃ for 1.0 min,and 72 ℃ for 1.5 min;and finally 72 ℃ for 10.0 min. Results of PAGE showed that clarity and amount of information increased with the new procedure. Thus,this superior method could be used for research on S. matsudana.[Ch,4 fig. 2 tab. 12 ref.]

WANG Hong-ling, ZHANG Jue, JIAO Zhong-yi, SHI De-zong. Establishing and optimizing the SRAP-PCR system for Salix matsudana[J]. Journal of Zhejiang A&F University, 2010, 27(6): 945-949. doi: 10.11833/j.issn.2095-0756.2010.06.024
Citation: WANG Hong-ling, ZHANG Jue, JIAO Zhong-yi, SHI De-zong. Establishing and optimizing the SRAP-PCR system for Salix matsudana[J]. Journal of Zhejiang A&F University, 2010, 27(6): 945-949. doi: 10.11833/j.issn.2095-0756.2010.06.024

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