N4-Methyl-dCTP
目录号 : GB20062N4-Methyl-dCTP（N4me-dCTP）是脱氧胞苷5'-三磷酸（dCTP）的类似物，应用于DNA或RNA的PCR合成。
Cas No.:149674-24-8
Sample solution is provided at 25 µL, 10mM.
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N4-Methyl-dCTP (N4me-dCTP) is an analog of deoxycytidine 5'-triphosphate (dCTP) that is applied in DNA or RNA synthesis with PCR[1]. N4-Methyl-dCTP naturally exists in some bacteria[2]. N4-Methyl-dCTP (200µM) improves the output of DNA synthesis at GC-rich region [1], and N4-Methyl-dCTP shows better acceptance by polymerase in SNP detection[3] due to overcoming strong secondary structure that interferes with primer annealing[4].
References:
[1] Flores-Juárez, Cyntia R et al. “PCR amplification of GC-rich DNA regions using the nucleotide analog N4-methyl-2'-deoxycytidine 5'-triphosphate.” *BioTechniques* vol. 61,4 175-182. 1 Oct. 2016, doi:10.2144/000114457
[2] Ehrlich, M et al. “DNA methylation in thermophilic bacteria: N4-methylcytosine, 5-methylcytosine, and N6-methyladenine.” *Nucleic acids research* vol. 13,4 (1985): 1399-412. doi:10.1093/nar/13.4.1399
[3] Yang, Zunyi et al. “Eliminating primer dimers and improving SNP detection using self-avoiding molecular recognition systems.” *Biology methods & protocols* vol. 5,1 bpaa004. 10 Feb. 2020, doi:10.1093/biomethods/bpaa004
[4]Swenson, Stephanie J, and Birgit Gemeinholzer. “Testing the effect of pollen exine rupture on metabarcoding with Illumina sequencing.” *PloS one* vol. 16,2 e0245611. 2 Feb. 2021, doi:10.1371/journal.pone.0245611
N4-Methyl-dCTP（N4me-dCTP）是脱氧胞苷5'-三磷酸（dCTP）的类似物，应用于DNA或RNA的PCR合成[1]。N4-Methyl-dCTP天然存在于某些细菌中。N4-Methyl-dCTP (200µM)提高了富GC区的DNA合成产量[1]，并且由于克服了干扰引物退火的强二级结构[4]，N4-Methyl-dCTP在SNP检测中被聚合酶更好地接受[3]。
用核苷酸类似物N4-Methyl-dCTP扩增富gcDNA区域[1]
1.引物设计:
使用Integrated DNA Technologies的PrimerQuest程序进行引物设计。选择在序列中含有最少鸟嘌呤的引物，以最大限度地降低含有N4meC的DNA模板的引物杂交温度。
2.PCR过程:
2.1制备PCR混合物：30ng人类基因组DNA；每种引物0.5µM；PCR缓冲液(50µM KCl; 10µM Tris-HCl; pH = 8.3);每种脱氧核糖核苷酸（dATP, dGTP, dTTP和dCTP）各200µM，或在修饰扩增子的情况下，用200µM的N4me-dCTP代替dCTP；1.5U HotStart Taq DNA聚合酶；2.5µM MgCl2;SYBR Green I的终浓度为0.4×。
2.2循环方案：95℃ 1min；然后进行45次95°C 30s的循环；62°C, 64°C, 或66°C 30s, 72°C 60s；最后在72°C下延长4min。
本实验方案仅供参考，需要根据实际需求进行修改
[1]Flores-Juárez, Cyntia R et al. “PCR amplification of GC-rich DNA regions using the nucleotide analog N4-methyl-2'-deoxycytidine 5'-triphosphate.” BioTechniques vol. 61,4 175-182. 1 Oct. 2016, doi:10.2144/000114457
| Cas No. | 149674-24-8 | SDF | Download SDF |
| 分子式 | C10H18N3O13P3 (free acid) | 分子量 | 481.1 (free acid) |
| 溶解度 | 储存条件 | Store at -20°C or below | |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.0786 mL | 10.3928 mL | 20.7857 mL |
| 5 mM | 0.4157 mL | 2.0786 mL | 4.1571 mL |
| 10 mM | 0.2079 mL | 1.0393 mL | 2.0786 mL |
Quality Control & SDS
- View current batch:
- Purity: >98.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet