快速和经济高效的水稻叶片DNA提取方法的比较评价

S. Nihad, Amirul Kabir, M. Rashid, Omma Honey, Mohammad Khan, Mohammad Latif
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引用次数: 1

摘要

水稻叶片DNA提取技术发展较早,但为了快速开展分子研究,仍在寻找一种快速、经济的DNA提取方法。对提取缓冲液(E1)、k -乙酸酯(E2)和两种改良CTAB (E3和E4)为基础的DNA提取方法进行了DNA浓度、提取DNA纯度、每个样品所需时间和成本的测试。此外,混合机(Mi;电法)和研钵和杵(M;采用手工方法)对四种方法的性能进行了测试。MiE1 (266.61 ng/µL)、MiE3 (260.58 ng/µL)和MiE4 (245.49 ng/µL)方法的DNA浓度差异不显著,但与MiE2 (137.13 ng/µL)方法差异显著。ME2所需时间最长(59分钟),其次是ME1(55分钟)、ME3(40分钟)和ME4(21分钟)。在成本方面,MiE3和MiE4的成本最高(356.39塔卡(4.19美元)/100个样品),而MiE2的成本最低(101.81塔卡(1.20美元)/100个样品)。纯度-1 (260/280)(PU-1)的MiE2与其他方法的MiE2差异不显著,纯度-2 (260/230)(PU-2)的MiE2与其他方法的MiE2差异显著。通过研钵和杵检测,ME1的DNA浓度最高(382.68 ng/µL),与ME2、ME3和ME4差异显著。ME2法所需时间最多(59 min),其次为ME1法(55 min)、ME3法(40 min)和ME4法(21 min)。ME3和ME4的成本最高(356.39塔卡(4.19美元)/100个样品),其次是ME2(121.83塔卡(1.43美元)/100个样品)和ME1(101.81塔卡(1.20美元)/100个样品)。不同方法间PU-1差异不显著,但PU-2、ME2与其他方法差异显著。基于CTAB的方法是提取DNA最好、最快速的方法,但成本较高,而其他两种方法较便宜,但耗时较长。
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Comparative assessment of quick and cost-efficient quality DNA extraction methods from rice leaves
DNA extraction from rice leaves was developed earlier but a quick and cost-efficient DNA extraction method is still in search to conduct molecular research swiftly. Four DNA extraction methods i.e., extraction buffer (E1), K-acetate (E2) and two modified CTAB (E3 and E4) based protocols were tested for DNA concentration, purity of the extracted DNA, time and cost required per sample. Moreover, a Mixer mill (Mi; electrical method) and a Mortar and pestle (M; manual method) were used to test the performance of the four methods. DNA concentrations are insignificant among the methods MiE1 (266.61 ng/µL), MiE3 (260.58 ng/µL) and MiE4 (245.49 ng/µL) but these are all significantly different from the method MiE2 (137.13 ng/µL). ME2 required the maximum time (59 minutes), followed by ME1 (55 minutes), ME3 (40 minutes), and ME4 (21 minutes). In terms of cost, MiE3 and MiE4 required the highest (356.39 taka (4.19 dollars)/100 samples) amount of cost, whereas the lowest price (101.81 taka (1.20 dollars)/100 samples) was required in MiE2. For Purity-1 (260/280) (PU-1), no significant difference was found among the methods, but for Purity-2 (260/230) (PU-2), MiE2 significantly differed from the other methods. Through the mortar and pestle, the highest DNA concentration was found in ME1 (382.68 ng/µL) which significantly differed from ME2, ME3 and ME4. The highest times required in ME2 (59 min) followed by ME1 (55 min), ME3 (40 min) and ME4 (21 min) methods. The highest costs (356.39 taka (4.19 dollars)/100 samples) were required for ME3 and ME4 methods followed by ME2 (121.83 taka (1.43 dollars)/100 samples) and ME1 (101.81 taka (1.20 dollars)/100 samples). No significant difference was found among the methods for PU-1 but for PU-2, ME2 significantly differed from other methods. CTAB-based methods are the best and quick for DNA extraction but these methods are costly, whereas the other two methods are cheaper but they are time-consuming compared to CTAB.
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