Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing |
| |
| Authors: | Qi WANG Xiaoxia SHEN Tian QIU Wei WU Lin LI Zhi'an WANG Huixia SHOU |
| |
| Institution: | 1. State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310058, China ; 2. Zhejiang Institute of Chinese Medicine, Hangzhou 310023, China ; 3. Shanghai YouLong Biotech Co., Ltd., Shanghai 200063, China |
| |
| Abstract: | Nucleic acids in plant tissue lysates can be captured quickly by a cellulose filter paper and prepared for amplification after a quick purification. In this study, a published filter paper strip method was modified by sticking the filter paper on a polyvinyl chloride resin (PVC) sheet. This modified method is named EZ-D, for EASY DNA extraction. Compared with the original cetyl trimethylammonium bromide (CTAB) method, DNA extracted by EZ-D is more efficient in polymerase chain reaction (PCR) amplification due to the more stable performance of the EZ-D stick. The EZ-D method is also faster, easier, and cheaper. PCR analyses showed that DNA extracted from several types of plant tissues by EZ-D was appropriate for specific identification of biological samples. A regular PCR reaction can detect the EZ-D-extracted DNA template at concentration as low as 0.1 ng/μL. Evaluation of the EZ-D showed that DNA extracts could be successfully amplified by PCR reaction for DNA fragments up to 3000 bp in length and up to 80% in GC content. EZ-D was successfully used for DNA extraction from a variety of plant species and plant tissues. Moreover, when EZ-D was combined with the loop-mediated isothermal amplification (LAMP) method, DNA identification of biological samples could be achieved without the need for specialized equipment. As an optimized DNA purification method, EZ-D shows great advantages in application and can be used widely in laboratories where equipment is limited and rapid results are required. |
| |
| Keywords: | DNA extraction Cellulose filter paper Field test Genetically modified (GM) crops Chinese medicine ultra-fine powder Rapid molecular identification |
| 本文献已被 CNKI 维普 等数据库收录！ |
|
