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Journal of Zhejiang University SCIENCE B
ISSN 1673-1581(Print), 1862-1783(Online), Monthly
2021 Vol.22 No.2 P.99-111
Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing
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/?L. 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.
Key words: DNA extraction; Cellulose filter paper; Field test; Genetically modified (GM) crops; Chinese medicine ultra-fine powder; Rapid molecular identification
1华中科技大学数字制造装备与技术国家重点实验室,中国武汉市,430074
2广东思谷智能技术有限公司,中国东莞市,523808
摘要:本文展示了一种可潜在帮助单侧膝受伤患者正常行走的传感引导步态同步下肢外骨骼系统。外骨骼能够减轻人体体重对受伤膝下肢的负载,并维持与健康侧下肢行走步态摆动相同步。传感引导步态同步系统集成了人体传感网络,它能感知健康侧下肢的运动步态。基于测量的关节角度轨迹引导,安装电机的髋关节在行走中提起腿杆,并将膝受伤步态和健康步态以半周期延时进行同步。实验验证了下肢外骨骼的效果。本文比较了健康腿和膝受伤腿的测量关节角度轨迹、仿真的膝受力、人机交互力等方面,结果说明髋关节安装电机受控制的下肢外骨骼能够将受伤腿和体重支撑外骨骼的融合步态与健康腿步态进行同步。
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DOI:
10.1631/jzus.B2000465
CLC number:
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On-line Access:
2021-02-07
Received:
2020-08-10
Revision Accepted:
2020-10-08
Crosschecked:
2021-01-06