QuestTaq™ PreMix is supplied as a convenient 2X concentrated “master mix” containing all the reagents (i.e., dNTPs, MgCl2, and enhancers) necessary for robust PCR with little or no by-product formation. The QuestTaq™ PreMix has been optimized for the non-biased amplification of cystosine, 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC), and glucosyl-5-hydroxymethylctosine (g5hmC) containing DNA, ensuring high yield amplification across a wide range of templates. The QuestTaq™ PreMix differs from QuestTaq™ qPCR PreMix (Cat. Nos. E2052 & E2053 from Zymo Research) in that it excludes SYTO® 9 dye from the PreMix solution, making it compatible with real-time and quantitative PCR with fluorescent dyes of the researcher’s choosing.
| Storage |
QuestTaq™ PreMix should be stored between -20 °C to -80 °C for maximum performance. Performance is guaranteed for 1 year from the time of receipt. |
| Unit Definition |
One unit (U) enzyme of QuestTaq™ DNA polymerase is defined as the amount of enzyme required for the incorporation of 10 nmol dNTPs into an acid-insoluble form in 30 minutes at 72°C. |
| Enzyme Concentration |
Reaction conditions at 1X (20 µl total volume) will contain 2 units of QuestTaq™ DNA polymerase. |
QuestTaq™ polymerase consistently yields robust amplicons from DNA templates having modified/unmodified cytosines. The figure shows the level (intensity) of an ~900 bp product generated from DNA templates using QuestTaq™ PreMix or the polymerases from Suppliers X and Y. In each case, PCR products were separated in a 0.8% (w/v) agarose/TAE/EtBr gel. Lanes correspond to amplicons from template DNA containing: unmodified cytosine (c), 5-methylcytosine (m), 5-hydroxymethylcytosine (h), or glucosyl-5-hydroxymethylcytosine (g). (M) is a 1 kb DNA Marker (Zymo Research, Cat. No. M5006-50).
Premixed reagents for one-tube PCR
Ideal for robust, non-biased amplification of 5mC, 5hmC, and g5hmC modified DNA
Ideal for real-time, quantitative, and end-point analyses
Compatible with a range of fluorescent dyes for use in real-time PCR
| QuestTaq™ PreMix |
50 rxns. |
E2050 |
 |
| QuestTaq™ PreMix |
200 rxns. |
E2051 |
|
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关于5-hmC
5-hmC是近年来在动物组织中发现的，由胞嘧啶修饰而来。5-hmC在表观遗传学上的功能可能与5-甲基化胞嘧啶(5-mC)不同。尽管到现在为止还不确知其功能，有研究者猜测它在调控基因的表达与关闭过程中起着重要的作用。5-mC的发现让我们不得不重新评估DNA甲基信息，也不得不监测人类的健康组织和病理组织之间5-mC相对分布的差异。在EPI公司的MethylFlash技术之前，我们还没有发现任何直接的常规方法来检测5- hmC,以及区分5-hmC和5-mC
5-hmC 和 5-mC的区别
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