HotStart™ 2X Probe qPCR Master Mix
![mRNA synthesis](/media/diy/images/page/figure1-mrna.png)
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
![Tyramine Signal Amplification (TSA)](/media/diy/images/page/figure2-01.png)
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
![screening library](/media/diy/images/page/figure3-01.png)
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
![Cell Counting Kit-8 (CCK-8)](/media/diy/images/page/CCK-8.jpg)
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
![SYBR Safe DNA Gel Stain](/media/diy/images/page/SYBR Safe DNA Gel Stain.png)
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
![Inhibitor Cocktails](/media/diy/images/page/Inhibitor Cocktails.jpg)
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
定量PCR(qPCR,又称Real-time PCR)是一种非常通用的精确分析基因表达的技术。按原理不同可分为染料法和探针法两类。常用的探针包括Taqman探针、分子信标、双杂交探针、MGB探针等。其中以TaqMan探针最为经典,使用最为广泛。Taqman探针能与目标序列特异性杂交,其5’ 端标记荧光基团,3’ 端标记淬灭基团。当探针完整时,由于荧光共振能量转移(FRET),探针上的荧光基团被淬灭基团淬灭。当引物介导的延伸反应到达探针位置时,Taq酶的5' → 3'外切酶活性会导致结合在目标序列上的探针逐渐被水解,5’ 端的荧光基团和3’ 端的淬灭基团分离,使它们的间距超过10 nm,超出荧光共振能量转移的范围,荧光基团此时在合适激发光的作用下,发出荧光。每经过一个PCR循环,就会有更多的荧光基团被释放,荧光强度与新合成的目标片段数量成正比,从而实现定量检测。探针5'端常用的荧光标记有FAM、HEX、TET、JOE、NED、Cy3、TAMRA,、Cy5、Cal Fluor Gold540、Cal Fluor Orange560等等,3'端标记常用BHQ1、BHQ2、BHQ3、TAMRA、Dabcyl、NFQ-MGB等淬灭基团,实际设计探针时,请根据qPCR仪的型号,需要检测的目的基因的不同,实验类型的不同来进行设计。
HotStart™ 2X Probe qPCR Master Mix是基于探针法实时荧光定量PCR的高品质预混液。核心组分Taq DNA Polymerase是基于抗体法修饰,经过升级改造提升模板亲和能力的新一代热启动DNA聚合酶,提高了对于血液、血清IgG、腐殖酸等PCR抑制成分的抵抗性,能够实现便捷高效的热启动。本产品特别适合用于低丰度或高特异性的目的基因的定量或定性检测。
本产品是一款2X预混液,只需额外加入引物、探针、模板即可,使用方便,同时兼容快速程序,缩短检测时间,可在宽广的动态范围内实现准确的,重复性高的目的基因定量。本产品可用于多重qPCR, 最多支持四重qPCR。
Components | 5 mL 1000 rxn with 10 μL reaction 500 rxn with 20 μL reaction 200 rxn with 50 μL reaction |
25 mL 5000 rxn with 10 μL reaction 2500 rxn with 20 μL reaction 1000 rxn with 50 μL reaction |
50 mL 10000 rxn with 10 μL reaction 5000 rxn with 20 μL reaction 2000 rxn with 50 μL reaction |
HotStart™ 2X Probe qPCR Master Mix | 1 mL X 5 | 1 mL X 25 | 5 mL X 10 |
50X ROX Reference Dye (low concentration) | 0.2 mL | 1 mL | 1 mL X 2 |
50X ROX Reference Dye (high concentration) | 0.2 mL | 1 mL | 1 mL X 2 |
Store all components at -20°C and protect from light for 12-24 months. Avoid repeated freeze/thaw cycles as possible. |