东北林业大学学报
東北林業大學學報
동북임업대학학보
JOURNAL OF NORTHEAST FORESTRY UNIVERSITY
2009年
11期
13-16
,共4页
孟宪婷%迟德富%王秀华%夏德安
孟憲婷%遲德富%王秀華%夏德安
맹헌정%지덕부%왕수화%하덕안
水曲柳(Fraxinus mandshurica Rupr.)%SRAP-PCR%正交设计%单因子试验
水麯柳(Fraxinus mandshurica Rupr.)%SRAP-PCR%正交設計%單因子試驗
수곡류(Fraxinus mandshurica Rupr.)%SRAP-PCR%정교설계%단인자시험
Fraxinus mandshurica%SRAP-PCR%Orthogonal design%Single factor test
以水曲柳叶片DNA为模板,利用SRAP(相关序列多态性)技术及L_(16)(4~5)正交试验和单因子试验方法,分析了不同浓度的DNA模板、Mg~(2+)、dNTP、引物、TaqDNA聚合酶对扩增结果的影响,最终确立的PCR最佳反应系统为:20μL体系中,2×PCR buffer、DNA模板50～100ng,Mg~(2+) 的浓度2.0mmol/L,dNTP0.15mmol/L,引物0.30μmol/L,TaqDNA聚合酶1.0U;最佳退火温度50℃.在此反应系统下,所扩增谱带清晰、稳定、多态性高.
以水麯柳葉片DNA為模闆,利用SRAP(相關序列多態性)技術及L_(16)(4~5)正交試驗和單因子試驗方法,分析瞭不同濃度的DNA模闆、Mg~(2+)、dNTP、引物、TaqDNA聚閤酶對擴增結果的影響,最終確立的PCR最佳反應繫統為:20μL體繫中,2×PCR buffer、DNA模闆50～100ng,Mg~(2+) 的濃度2.0mmol/L,dNTP0.15mmol/L,引物0.30μmol/L,TaqDNA聚閤酶1.0U;最佳退火溫度50℃.在此反應繫統下,所擴增譜帶清晰、穩定、多態性高.
이수곡류협편DNA위모판,이용SRAP(상관서렬다태성)기술급L_(16)(4~5)정교시험화단인자시험방법,분석료불동농도적DNA모판、Mg~(2+)、dNTP、인물、TaqDNA취합매대확증결과적영향,최종학립적PCR최가반응계통위:20μL체계중,2×PCR buffer、DNA모판50～100ng,Mg~(2+) 적농도2.0mmol/L,dNTP0.15mmol/L,인물0.30μmol/L,TaqDNA취합매1.0U;최가퇴화온도50℃.재차반응계통하,소확증보대청석、은정、다태성고.
Effects of different concentrations of DNA template, Mg~(2+) , dNTP, primer, Taq DNA polymerase on amplification re-sult were studied by single factor test, orthogonal design of L_(16)(4~5) and Sequence-related amplified polymorphism marker (SRAP) technique, using the genomic DNA extracted from Fraxinus mandshurica Rupr as template. The optimal PCR system was obtained as 2 × PCR buffer, Mg~(2+) 2.0 mmol/L, dNTPs 0.15 mmol/L, primer 0. 30 μmol/L, Taq DNA poly-merase 1.0 U, and genomic DNA template 50～100 ng in 20 μL reaction system. And the optimal annealing temperature was 50 degrees C. Clear, stable and abundant polymorphic bands were obtained under the optimal reaction system.
