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Comparison of multiplex PCR kits for SCoT and SRAP genotyping in plants
Plant Genetic Resources ( IF 1.2 ) Pub Date : 2021-02-24 , DOI: 10.1017/s1479262121000046
Ponsit Sathapondecha , Lathapol Boonsermsukchareon , Sukhuman Whankeaw

Taq polymerase system is one of the key success factors for genotyping by using semi-random marker types. Start codon targeted (SCoT) and sequence-related amplified polymorphism (SRAP) are semi-random markers that amplify more than one target at a time, like in multiplex PCR. Here, we compared the performance of five multiplex PCR kits (QIAGEN®, Thermo Scientific™, New England Biolabs, KAPA Biosystem and iNtRON Biotechnology) and Taq DNA polymerase from Vivantis as a representative common Taq polymerase. The PCR using three SCoT and three SRAP markers was performed in six plants, which were different in genome size, including three monocotyledons (rice, oil palm and onion) and three dicotyledons (holy basil, chilli pepper and eggplant). The overall results demonstrated consistent high performance of the Taq polymerase system with the multiplex PCR kit from KAPA Biosystem. Considered by plant, the multiplex PCR kit from KAPA Biosystem showed the best performance in holy basil, eggplant, oil palm and chilli pepper, whereas in rice, the Taq DNA polymerase from Vivantis generated better results than the KAPA Biosystem. In onion, multiplex PCR kit from Thermo Scientific™ showed the best performance among those tested, even if it did not perform well in the other plans. Different Taq polymerase systems might generate different band patterns, therefore using the same Taq polymerase system within one experimental dataset is recommended. The multiplex PCR kit from KAPA Biosystem is the best starting choice.

中文翻译:

用于植物 SCoT 和 SRAP 基因分型的多重 PCR 试剂盒的比较

塔克聚合酶系统是使用半随机标记类型进行基因分型的关键成功因素之一。起始密码子靶向 (SCoT) 和序列相关扩增多态性 (SRAP) 是半随机标记,可一次扩增多个靶标,就像在多重 PCR 中一样。在这里,我们比较了五种多重 PCR 试剂盒(QIAGEN®、Thermo Scientific™、New England Biolabs、KAPA Biosystem 和 iNtRON Biotechnology)和塔克来自 Vivantis 的 DNA 聚合酶作为代表常见塔克聚合酶。使用三个 SCoT 和三个 SRAP 标记的 PCR 在六种基因组大小不同的植物中进行,包括三种单子叶植物(水稻、油棕和洋葱)和三种双子叶植物(圣罗勒、辣椒和茄子)。总体结果表明该系统始终如一的高性能塔克聚合酶系统与来自 KAPA Biosystem 的多重 PCR 试剂盒。从植物方面考虑,KAPA Biosystem 的多重 PCR 试剂盒在圣罗勒、茄子、油棕和辣椒中表现出最佳性能,而在水稻中,塔克Vivantis 的 DNA 聚合酶产生了比 KAPA 生物系统更好的结果。在洋葱中,Thermo Scientific™ 的多重 PCR 试剂盒在测试中表现出最好的性能,即使它在其他计划中表现不佳。不同的塔克聚合酶系统可能会产生不同的条带模式,因此使用相同的塔克建议使用一个实验数据集中的聚合酶系统。KAPA Biosystem 的多重 PCR 试剂盒是最佳的起始选择。
更新日期:2021-02-24
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