Optimization on SSR-PCR reaction system of Chinese firChinese Full Text
XU Yang;CHEN Jinhui;WANG Ying;ZHAO Yaqi;WANG Xinmin;ZHENG Renhua;SHI Jisen;Key Laboratory of Forest Genetics and Biotechnology,Ministry of Education,Nanjing Forestry University;
Abstract: It’s important to optimize the SSR-PCR（ Polymerase Chain Reaction） reaction system firstly when doing research with SSR markers in Chinese fir. The orthogonal design of L 16（ 4）5was used to optimize the SSR-PCR system for Chinese fir with four levels and five factors,namely,Mg2 +,dNTPs,primer,DNA polymerase and DNA template. The results of PCR were evaluated by variance and intuitive analysis. Then,we respectively determined the concentration of Mg2 +, dNTPs,primer and annealing temperature using single factor experiment design. The results showed that the optimum reaction system was 20 μL reaction solution,containing Mg2 +1. 0mmol / L,dNTPs 0. 15 mmol / L,primer 0. 25 μmol / L,Taq polymerase 0. 05 U / μL,DNA template 30 ng,2 μL 10 × PCR buffer and adding ddH 2 O to 20 μL,and the annealing temperature was the 63- 53℃. To verify the reaction system,we used 4 pairs of EST-SSR markers to amplify 10 F 1 Chinese fir plantlets and their parents,and we got clear and stable bands,which showed that the SSR-PCR system was suitable further SSR research in Chinese fir.
- DOI：
10.13360/j.issn.1000-8101.2014.01.004
- Series：
- Subject：
- Classification Code：
S791.27
- Mobile Reading
Read on your phone instantly
Step 1Scan QR Codes
"Mobile CNKI-CNKI Express" App
Step 2Open“CNKI Express”
and click the scan icon in the upper left corner of the homepage.
Step 3
Scan QR Codes
Read this article on your phone.
- HTML
- CAJ Download
- PDF Download
Download the mobile appuse the app to scan this coderead the article.
Tips: Please download CAJViewer to view CAJ format full text.
Download: 281 Page: 15-20 Pagecount: 6 Size: 543K
Citation Network
Related Literature
- Similar Article
- Reader Recommendation
- Associated Author