Establishing and Optimizing AFLP Amplification Reaction System of Shiraia Bambusicola
Bing Yang, Yi-Meng Chen, Yong-Xiang Liu, Zuo-Yi Liu, De-Qun Zhou
Available Online June 2014.
- https://doi.org/10.2991/icmsa-15.2015.33How to use a DOI?
- Shiraia Bambusicola, AFLP, System Optimizing, Polymorphic Primer.
- DNA of Sharaia bambuiscola was extracted by the improved method of CTAB. The several key factors affecting the effect of DNA digestion and the PCR selective amplification such as the time of DNA digestion, the times of the per-amplified dilute products, the mount of the selective amplification primer, Taq concentration and dNTPs concentration were optimized and trialed with establishment of an optimized AFLP reaction system of S. bambusicola. The best time of digestion DNA with double endonucleases (EcoR and Mse ) was 3h. The optimized selection amplification system was 25µl reaction mix containing 2.5µl the 20 times of the per-amplified dilute products, 2.5µL 10× buffer(with Mg2+), 10mmol/L primer each 1.5µL, 2.5U•µL-1 Taq polymerase 0.7µL, 10mmol/L dNTP 0.5µL, and 15.8µL ddH2O. Stable and clean DNA finger print can be obtained and 3 pairs of AFLP primers with good genetic diversity were selected according to the optimized reaction system. The results will be an effective protocol for further studying the genetic stucture and differentiation, phylogenetic trees, host specificity and artificial cultivation of Shiraia bambusicola population.
- Open Access
- This is an open access article distributed under the CC BY-NC license.
Cite this article
TY - CONF AU - Bing Yang AU - Yi-Meng Chen AU - Yong-Xiang Liu AU - Zuo-Yi Liu AU - De-Qun Zhou PY - 2014/06 DA - 2014/06 TI - Establishing and Optimizing AFLP Amplification Reaction System of Shiraia Bambusicola BT - 2015 International Conference on Material Science and Applications (icmsa-15) PB - Atlantis Press SP - 173 EP - 180 SN - 2352-541X UR - https://doi.org/10.2991/icmsa-15.2015.33 DO - https://doi.org/10.2991/icmsa-15.2015.33 ID - Yang2014/06 ER -