An Optimized Preparation Method to Obtain High-quality RNA from the Buds of Sunflowers
- DOI
- 10.2991/emcpe-16.2016.18How to use a DOI?
- Keywords
- Sunflower Buds; RNA Extraction; Polyphenol; Polysaccharide
- Abstract
In an attempt to isolate high-quality, total RNA from flower buds of sunflowers (Helianthus annuus). We have already improved the SDS method. After adding liquid nitrogen, we have to grind the material down. After that we add protein -DNA precipitation, wash it with the addition of different kinds of alcohol, such as 75% alcohol and precipitation. Then we add DEPC water and guanidine hydrochloride blend, LiCl overnight precipitation,and then wash it off with 75% alcohol, finally make it dissolved in 15 æL DEPC water which collocated in minus 80 environment for storage. The total RNA is precipitated with 1/10 volume of NaAc and 2 volumes of absolute ethanol to prevent contamination by polysaccharides. These experimental results show that the total RNA extracted by this method is of high purity and integrity, and the electrophoretic bands are clear. D260/D230, D260/D280 and RNA (g/100mg) are 2.05, 2.11 and 9.52, respectively. Total RNA can amplify the target gene fragment by PT-PCR. This improved method can be used to extract the high-quality RNA from the sunflower buds of the space mutation, in order to study the molecular mechanism of the next step.
- Copyright
- © 2016, the Authors. Published by Atlantis Press.
- Open Access
- This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).
Cite this article
TY - CONF AU - Hao Feng AU - XiBao Ma AU - Jun Yang PY - 2016/08 DA - 2016/08 TI - An Optimized Preparation Method to Obtain High-quality RNA from the Buds of Sunflowers BT - Proceedings of the 2016 5th International Conference on Environment, Materials, Chemistry and Power Electronics PB - Atlantis Press SP - 78 EP - 85 SN - 2352-5401 UR - https://doi.org/10.2991/emcpe-16.2016.18 DO - 10.2991/emcpe-16.2016.18 ID - Feng2016/08 ER -