AmpErase™ Uracil N-Glycosylase (UNG)
AmpErase™ Uracil N-Glycosylase (UNG)
Applied Biosystems™

AmpErase™ Uracil N-Glycosylase (UNG)

AmpErase™ Uracil N-Glycosylase (UNG), part of the GeneAmp™ PCR Carry-over Prevention Kit, is a 26 kDa ultrapure, recombinant enzyme encoded詳細を見る
製品番号(カタログ番号)数量
N8080096100 units
製品番号(カタログ番号) N8080096
価格(JPY)
43,300
Each
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数量:
100 units
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AmpErase™ Uracil N-Glycosylase (UNG), part of the GeneAmp™ PCR Carry-over Prevention Kit, is a 26 kDa ultrapure, recombinant enzyme encoded by the E. coli uracil N-glycosylase gene, which has been inserted into an E. coli host to direct the expression of the wild type form of the enzyme.The enzyme removes any uracil incorporated into single- or double-stranded DNA.

GeneAmp™ PCR Carry-over Prevention Kit
The GeneAmp™ PCR Carry-over Prevention Kit (available separately) provides reagents for a simple yet powerful method of ensuring that PCR products cannot be reamplified in subsequent PCR amplifications, thereby preventing false positive results.このキットの特長:

•Enzymatic method stops PCR carryover contamination, which prevents false positive results
• Designed to degrade PCR products from previous PCR amplifications without degrading native nucleic acid templates, which improves amplification
• No interference with any PCR or real-time PCR application
• Optimized for use with GeneAmp™ PCR core reagents and GeneAmp™ instrument systems

Eliminate PCR carryover contamination
The kit uses an enzymatic reaction analogous to the restriction-modification and excision-repair systems of cells to specifically degrade PCR products from previous PCR amplifications in which dUTP has been incorporated, without degrading native nucleic acid templates.The method used to render PCR products susceptible to degradation involves substituting dUTP for dTTP in the PCR mixture, and pretreating all subsequent PCR mixtures with AmpErase™ Uracil N-glycosylase (UNG) prior to PCR amplification.Products from PCR amplification contain uracil, and are readily distinguishable from native thymidine-containing DNA templates.Products from previous PCR amplifications are eliminated by excising uracil residues using UNG, and degrading the resulting abasic polynucleotide with heat.Although UNG is active on single- and double-stranded dU-containing DNA, ribouracil residues in RNA and dUTP itself are not substrates for UNG.AmpliTaq™ DNA Polymerase, AmpliTaq Gold™ DNA Polymerase, and the other components of the PCR mixture remain intact during UNG treatment, providing PCR amplification free of PCR product carryover.The dU-containing PCR product behaves like dT-containing DNA in blotting, cloning, sequencing, and most other post-PCR analyses.
研究用にのみ使用できます。診断用には使用いただけません。
仕様
ポリメラーゼDNAポリメラーゼ
製品ラインAmpErase
製品タイプPCR反応用のウラシルN-グリコシラーゼ
数量100 units
出荷条件ドライアイス
対応可能対象100 PCR増幅
使用対象(アプリケーション)Standard PCR
PCR法qPCR, Standard PCR
Unit SizeEach
組成および保存条件
Contains AmpErase™ Uracil N-glycosylase (UNG) (100 units).各100 µLのPCR増幅100回分です。-20℃で保存

よくあるご質問(FAQ)

What is AmpErase UNG, and how does it work?

AmpErase UNG (Uracil N-glycosylase) is an enzyme utilized in a powerful method for elimination of carryover PCR products in Real-Time PCR. This method modifies PCR products such that in a new reaction, any residual products from previous PCR amplifications will be digested and prevented from amplifying, but the true DNA templates will be unaffected.

Here is how it works: During amplification, dUTP is substituted for dTTP, resulting in dUTP-containing amplicons. In subsequent reactions, a short pre-PCR incubation step will allow the AmpErase UNG to digest any dUTP containing DNA. Since AmpErase UNG is active on both single- and double-stranded dUTP-containing DNA, the procedure should work on dU-containing PCR products from standard or asymmetric PCR amplifications. However, uracil ribonucleotide residues in RNA, novel DNA containing dTTP, or cDNA containing dTTP will not be suitable substrates for UNG, so your templates will be unaffected.

Note that this is a proactive method to prevent contamination from future reactions, but will not help with a pre-existing contamination problem of standard dTTP-containing PCR products. That can only be remedied with thorough cleaning of lab surfaces, equipment and air filters.

I am using AmpErase Uracil N-glycosylase (UNG) and dUTP in my PCR reactions and would like to use the PCR product in a post-PCR application. Does the dUTP affect my ability to hybridize, sequence, clone, or digest the PCR product?

PCR products containing dUTP residues are generally equivalent to dT-containing PCR products as hybridization targets and as templates for dideoxy-terminated sequencing reactions. They can also work equivalently as targets for direct cloning as long as they are transferred into UNG-minus bacterial hosts.

The recognition of dU-containing DNA by restriction endonucleases has been studied, and depending on the specific endonuclease there may be no effect on enzymatic activity on the substitution of dU for dT (e.g., EcoR1 and BamH1), or the dU-containing DNA is cleaved more slowly than dT-containing DNA (e.g., Hpa1, HindII, and HindIII). The most accurate answer on the effects of substituting dU for dT are best determined using your own empirical studies.