ExoSAP-IT™ PCR 产物清洁纯化
Applied Biosystems™

ExoSAP-IT™ PCR 产物清洁纯化

ExoSAP-IT™ PCR 产物纯化的特点和优势• 保存 PCR 样品—短片段和长片段 PCR 产物的回收率为 100%•一管/一步法 PCR了解更多信息
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货号数量
78250.40.UL40 μL
货号 78250.40.UL
价格(CNY)
607.00
Each
数量:
40 μL
价格(CNY)
607.00
Each
ExoSAP-IT™ PCR 产物纯化的特点和优势
保存 PCR 样品—短片段和长片段 PCR 产物的回收率为 100%
一管/一步法 PCR 纯化—直接向 PCR 产物中加入 ExoSAP-IT 试剂
无需离心柱—节省时间和费用,同时提高得率
去除污染的引物和 dNTP—不干扰下游应用
可扩展—高通量纯化更经济
过程简单—自动;替代磁珠、过滤和板
离心柱法

ExoSAP-IT 试剂设计用于简单、快速的 PCR 纯化,用于下游应用,如 DNA 测序或单核苷酸多态性 (SNP) 分析。PCR 扩增完成时,产物中剩余的任何未消耗的 dNTP 和引物都会干扰这些方法。ExoSAP-IT 可去除这些污染物。

ExoSAP-IT 可直接添加到 PCR 产物中并在 37°C 下孵育 15 分钟(图 1)。PCR 处理后,ExoSAP-IT 只需加热至 80°C 15 分钟即可失活。

ExoSAP-IT 一步 PCR 纯化使用两种水解酶,Exonuclease I 和 Shrimp Alkaline Phosphatase (SAP),一起溶于特殊的配制缓冲液中,去除 PCR 产物中不需要的 dNTP 和引物。Exonuclease I 去除 PCR 中产生的残留单链引物和任何无关的单链 DNA。SAP 可去除 PCR 混合物中的剩余 dNTP。

快速PCR 产物清洁纯化方案
ExoSAP-IT 仅需一步移液和两次孵育。只需将 ExoSAP-IT 添加到 PCR 产物中,即可在 30 分钟内完成测序或 SNP 分析。

简单:一步
方法设计仅需较短的‘手动操作’时间。通过在单管或微量滴定板孔中使用 ExoSAP-IT 试剂,可以在一个简单的步骤中完成酶促去除多余的引物和未掺入的核苷酸。因此,只需简单的移液器转移,可同时手动或自动处理许多样品。

无样品损失
使用 ExoSAP-IT 试剂无需进行所有凝胶或柱纯化、沉淀、过滤、微珠和/或磁性分离。使用 ExoSAP-IT 的短和长 PCR 产物回收率为 100%(图 2)。

从 PCR 产物获得高质量数据
ExoSAP-IT 试剂可用作高效纯化方法,后续可进行荧光或放射性 DNA 测序、SNP 分析或任何其他需要不含多余核苷酸和引物的 PCR 产物的应用。

参考文献:
DUGAN, K. A., LAWRENCE, H. S., HARES, D. R., FISHER, C. L. AND BUDOWLE B. (2002) J. Forensic Sci 47, 811-818.
HANKE, M. AND WINK, M. (1994) BioTechniques 17, 858-860.

MU, J., DUAN, J., MAKOVA, K., JOY, D., HUYNH, C., BRANCH, O., LI, W. AND SU, X. (2002) Nature 418, 323-326.
SILVA, JR., W. A., COSTA, M. C. R., VALENTE, V., DE FREITAS SOUSA, J., PACÓ-LARSON, M. L., ESPREAFICO, E. M., CAMARGO, S. S., MONTEIRO, E., DE JESUS, A., HOLANDA, M. A., ZAGO, M. A., SIMPSON, A. J. G. AND NETO, E. D. (2001) BioTechniques 30, 537-542.
WERLE, E., SCNEIDER C., RENNER, M., VÖLKER, M. AND FIEHN, W. (1994) Nucleic Acids Res.22, 4354-4355.
仅供科研使用。不可用于诊断程序。
规格
描述ExoSAP-IT™ PCR Product Clean-up
适用于(应用)PCR 纯化
数量40 μL
测试时间15 分钟
产品规格离心柱
Unit SizeEach

常见问题解答 (FAQ)

Can ExoSAP-IT PCR cleanup reagents be used with automatic handling systems?

The HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup reagent is designed for automated handlers as it has a lower viscosity than standard ExoSAP-IT PCR Product Cleanup reagent, which contains some glycerol.

After purification with ExoSAP-IT PCR cleanup reagents, what is the recommended quantification method?

The Quant-iT PicoGreen dsDNA Assay Kit is recommended for quantifying PCR products that have been cleaned with ExoSAP-IT reagent. ExoSAP-IT PCR cleanup results in 100% recovery regardless of amplicon length, and if the PCR product was quantified prior to ExoSAP-IT cleanup, additional quantification is not necessary. We do not recommend using a spectrophotometer as degraded products will be included in the quantification.

Can ExoSAP-IT PCR cleanup reagents be used for removal of primers after first-strand cDNA synthesis?

For use with reverse transcription, ExoSAP-IT does not degrade the DNA-RNA hybrid. However, it will degrade single-stranded cDNA if a 3' -OH end is present.

Will the two tracking dyes used for visualization in the DreamTaq Hot Start Green PCR Master Mix interfere with the ExoSAP-IT PCR cleanup reagents?

The tracking dyes used in the DreamTaq Hot Start Green PCR Master Mix have not been shown to interfere with functionality of the ExoSAP-IT cleanup reagents.

Can ExoSAP-IT PCR cleanup reagents be used to cleanup PCR products amplified using High Fidelity (HiFi) DNA polymerases?

The proofreading function of HiFi DNA polymerases can cause partial degradation of PCR products during extended incubation at 80°C. ExoSAP-IT Express reagent is recommended for HiFi PCR cleanup because it only requires 1 minute at 80°C for inactivation.