Title

用正交設計優化荔枝RAPD反應體系

Translated Titles

Optimization of the RAPD Reaction System of Litchi chinensis Sonn. by Orthogonal Design

Authors

王家保(Jia-Bao Wang);劉志媛(Zhi-Yuan Liu);徐碧玉(Bi-Yu Xu);鄧穗生(Sui-Sheng Deng);杜中軍(Zhong-Jun Du);陳業淵(Ye-Yuan Chen)

Key Words

荔枝 ; RAPD ; 反應體系 ; 正交設計 ; Litchi ; RAPD ; Reaction system ; Orthogonal design

PublicationName

植物科學學報

Volume or Term/Year and Month of Publication

23卷4期(2005 / 08 / 01)

Page #

363 - 368

Content Language

簡體中文

Chinese Abstract

以改良CTAB法提取的荔枝基因組DNA為模板,應用L25(5^6)正交表,研究了Taq、Mg(上標 2+)、隨機引物、dNTPs和DNA模板5種RAPD反應組分濃度變化對擴增結果的影響,量化分析結果表明:正交設計可以應用於RAPD反應體系的建立。用這種方法建立的荔枝RAPD-PCR優化反應體系為:25μL反應體系中含1×Buffer、2.0 mmol/L Mg(上標 2+)、2.0 U Taq DNA聚合酶、0.15 mmol/L dNTPs、0.6μmol/L隨機引物、25ng DNA模板。

English Abstract

The L25(5^6) orthogonal diagram was applied to study the effects of different concentrations of Taq polymerase, Mg(superscript 2+), dNTPs, primer and DNA on litchi (Litchi chinensis Sonn.) RAPD-PCR results and to get a suitable PCR system. The results showed that the orthogonal design could be used to optimize the RAPD-PCR system. A suitable RAPD reaction system for litchi was developed, i. e. 1×Taq polymerase reaction buffer, 2.0 mmol/L Mg(superscript 2+) 2.0 U Taq DNA polymerase, 0.15 mmol/L dNTPs, 0.6 μmol / L primer and 25 ng genomic DNA templates with total 25 μL reaction solution.

Topic Category 生物農學 > 植物學