Key Difference – SYBR Green vs Taqman
SYBR Green和Taqman是使用两种用于检测或观察放大过程的方法real-time PCR. SYBR Green is a method based on intercalatingnucleic acidstaining dye while Taqman is a method based on hydrolysis probe. Both technologies are designed to generate fluorescence during the PCR, which allows real-time PCR machine to monitor the reaction in “real time”.使用称为SYBR绿色的荧光染料进行SYBR绿色方法,并通过将染料与产生的双链DNA结合来检测扩增。塔克曼使用双标记探针进行,并通过通过降解来检测探针的扩增Taq polymeraseand releases of the fluorophore.这是Sybr Green和Taqman之间的关键区别。
内容
1.Overview and Key Difference
2.什么是sybr绿色
3.What is Taqman
4.Side by Side Comparison – SYBR Green vs Taqman
5。概括
什么是sybr绿色?
SYBR Green is a fluorescent dye used to stain nucleic acids, especially double stranded DNA in Molecular Biology. SYBR Green method is used to quantify PCR products during the real-time PCR. Once it binds with DNA, the resulting DNA-dye complex absorbs blue light and emits intense green light. It happens due to the structural change that occurs in the dye molecule upon binding with double-stranded DNA. When PCR creates more and more DNA, more dye molecules bind with DNA, generating more fluorescence. Therefore, the fluorescence increases with the PCR product accumulation. Hence, the amount of PCR product can be quantitatively measured by the SYBR Green fluorescence detection.
SYBR绿色染料也可用于细胞仪和荧光显微镜中的DNA标记。溴化乙锭已被Sybr绿色成功取代,因为溴化乙锭是一种致癌染料,在凝胶电泳中DNA可视化期间具有处置问题。
SYBR绿色方法存在优势和缺点。该方法非常敏感,便宜且易于使用。但是,由于其与任何双链DNA结合的能力,非特异性结合可以导致PCR产物的过度定量。
What is Taqman?
Taqman是SYBR Green监视实时PCR过程的另一种方法。此方法取决于5' - 3'exonucleaseactivity of Taq polymerase enzyme to degrade theprobes在扩展新链和荧光团的释放期间。双标记探针用于此方法,它基于探针的水解。探针是荧光标记的DNA寡核苷酸,在5'端具有荧光记者分子(荧光团)和3'端的淬灭剂分子。它们被设计为在引物退火的另一侧与单链模板结合。TAQ聚合酶添加核苷酸to theprimerand extends the new strand towards the dual-labeled probes. Once the Taq polymerase meets the probe, exonuclease action of the Taq polymerase activates and degrades the probe. Once it completes the synthesis of the new strand, the probe is subjected to complete degradation and release the fluorophore. The release of fluorophore generates fluorescence. Fluorescent Quencher molecule efficiently quenches the emitted light and creates the output for quantification of the PCR product. The release of fluorophores and quantity of the PCR products are proportional. Hence, quantification can be easily done by the Taqman method.
Taqman方法用于实时PCR,定量gene expression, detection of genetic多态性, quantification of chromosomal DNA deletions, bacterial identification, verification of microarray analysis,SNP基因分型等
What is the difference between SYBR green and Taqman?
SYBR Green vs Taqman |
|
SYBR Green is based on DNA binding dye. | Taqman取决于杂交探针和TAQ聚合酶的5'至3'外切酶活性。 |
Fluorescently Labeled Probes | |
没有发出荧光ntly labeled probes are not required. | 需要双标记的探针。 |
多重基因分析 | |
It can not be used for multiplex gene targets. | 它可用于多重基因靶标。 |
成本 | |
这很便宜。 | This is more expensive. |
特异性 | |
This is less specific and binds with any double strand DNA | These are highly specific since probes detect the specific amplification products. |
Effectiveness | |
这是不太效果的。 | 这是非常有效的。 |
Application | |
这用于实时PCR,琼脂糖凝胶可视化,DNA标记等。 | 这用于实时PCR,基因表达的定量,遗传多态性的检测等。 |
概括– SYBR Green and Taqman
塔克曼and SYBR green are two methods employed in real-time PCR (quantitative PCR). Both methods enable the quantification of the PCR product efficiently and rely on the emission of the fluorescence. Taqman method uses dual-labeled probes for detection of the accumulated DNA while SYBR Green method uses a fluorescent dye. Both these methods also have different applications in molecular biology.
Reference:
1. Tajadini, Mohamad Hasan, Mojtaba Panjehpour, and Shaghayegh Haghjooy Javanmard. “Comparison of SYBR Green and TaqMan methods in quantitative real-time polymerase chain reaction analysis of four adenosine receptor subtypes.” Advanced Biomedical Research. Medknow Publications & Media Pvt Ltd, 2014. Web. 13 Mar. 2017.
2. “Real-time PCR basic principles.” Real Time PCR, Quantitative (qPCR), Primers & Mastermix : Primerdesign Ltd. N.p., n.d. Web. 13 Mar. 2017.
3.“ SYBR Green和其他实时PCR染料。”生物组合。N.P.,2010年4月5日。2017年3月14日
Image Courtesy:
1. “PCR with SYBR green” By –Ygonaar 23:09, 7 March 2006 (UTC) – It’s a graph create by Ygonaar with Power point, CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=619528
2.用户的“ Taqman”:脑损坏 - 自己的工作(公共领域)通过Commons Wikimedia
farzeena说
Thank you