Proceedings of the 2015 International Conference on Material Science and Applications

Establishing and Optimizing AFLP Amplification Reaction System of Shiraia Bambusicola

Authors
Bing Yang, Yi-Meng Chen, Yong-Xiang Liu, Zuo-Yi Liu, De-Qun Zhou
Corresponding Author
Bing Yang
Available Online June 2014.
DOI
https://doi.org/10.2991/icmsa-15.2015.33How to use a DOI?
Keywords
Shiraia Bambusicola, AFLP, System Optimizing, Polymorphic Primer.
Abstract
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.
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This is an open access article distributed under the CC BY-NC license.

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Proceedings
2015 International Conference on Material Science and Applications (icmsa-15)
Part of series
Advances in Physics Research
Publication Date
June 2014
ISBN
978-94-62520-75-2
ISSN
2352-541X
DOI
https://doi.org/10.2991/icmsa-15.2015.33How to use a DOI?
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  -