DescriptionViPrimePLUS Taq qPCR Green Master Mix II is next generation master mix designed for fast and easy realtime PCR reaction set up. The master mix is prepared in 2X concentrated solution and contains pure Taq DNA Polymerases, EvaGreen® dye, highest quality dNTPs, and buffer components at optimal concentrations. Taq DNA Polymerases in the master mix provide excellent results in reaction efficiency, correlation coefficient and slope. EvaGreen® dye in master mix is environmentally safe and highly stable which can be formulated with relative high dye concentration to maximize fluorescence signal without PCR inhibition.ViPrimePLUS Taq qPCR Green Master Mix II can be used to amplify any DNA template including genomic, cDNA and viral sequences. The formulation of qPCR green master mix can detect low copy number targets very specifically with high efficiency. The qPCR green master mix provides convenient and robust set up for quantitative real-time analysis of DNA samples.ViPrimePLUS Taq qPCR Green Master Mix II has several formulations optimized to be used with most of real-time PCR instruments. The improved sensitivity and consistency of ViPrimePLUS Taq qPCR Green Master Mix II in standard cycling conditions gives the industry leading performance in fast cycling conditions.
Applications
Features
- Ready-to-use real-time PCR reaction set up
- Rapid extension rate for early Ct values
- Good buffer system for excellent amplification efficiency
- Contain EvaGreen® dye – highest dye stability and safety
- High sensitivity detection
- Minimal PCR inhibition
- Compatible with most of the real-time PCR platforms
Component1.6ml aliquots of master mix
StorageStable at -20°C up to the expiry date stated. Store all components at -20°C upon arrival. Keep in aliquot to reduce freeze-thaw cycles.
Quality ControlAs part of the ISO9001:2008 quality assurance systems, each lot of ViPrimePLUS Taq qPCR Green Master Mix II has been tested against predetermined specifications to ensure consistent product quality and highest levels of performance and reliability.
Limitation Of UseFor research use only. Not recommended or intended for diagnosis of disease in humans or animals. Do not use internally or externally in humans or animals.
InstrumentsTo calibrate a real-time PCR reaction, various formulations of master mixes are available for most of the platforms.
| Product Description | Compatible Hardware |
| QLMM17ViPrimePLUS Taq qPCR Green Master Mix II (EvaGreen® Dye) | Biometra qTower, BioRad iCycler, BioRad IQ4, BioRad IQ5, Cepheid SmartCycler®, Eppendorf Mastercycler, Fluidigm BioMark™, Illumina Eco, MJ Chromo4, Opticon, PCRMax Eco™, Roche lightcycler® 480, lightcycler® LC96 and lightcycler® Nano Platforms, RotorGene, Roche Capillary Lightcycler 1.0-2.0, Stratagene MX MX4000P®, MX3000P®, MX3005®, Thermo PikoReal™ |
| QLMM17-LRViPrimePLUS Taq qPCR Green Master Mix II with Low ROX (EvaGreen® Dye) | Applied Biosystems 7500 and 7500 FAST platform, QuantStudio™, ViiA7 |
| QLMM17-RViPrimePLUS Taq qPCR Green Master Mix II with ROX (EvaGreen® Dye) | Applied Biosystems 7000, 7300, 7700, 7900 and 7900HT FAST platforms, GeneAmp® 5700, StepOne™, StepOne™ PLUS |
Ordering Information
| Catalog No | Description | Pack Size |
| QLMM17 | ViPrimePLUS Taq qPCR Green MasterMix II (EvaGreen® Dye) | 150 reactions |
| QLMM17-LR | ViPrimePLUS Taq qPCR Green MasterMix II with Low ROX (EvaGreen® Dye) | 150 reactions |
| QLMM17-R | ViPrimePLUS Taq qPCR Green MasterMix II with ROX (EvaGreen® Dye) | 150 reactions |
DownloadManual
ViPrimePLUS Taq qPCR Green MasterMix II (EvaGreen® Dye) ViPrimePLUS Taq qPCR Green MasterMix II with Low ROX (EvaGreen® Dye) ViPrimePLUS Taq qPCR Green MasterMix II with ROX (EvaGreen® Dye)
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accctcccgccccccccgtat
(互补碱基不写了）
那么一般来说,你就要导入以accctc结尾的引物使得增幅继续下去,具体方法和注意事项楼主可以看gee_an大侠的资料.
查了一下测序公司的网页,如果要同定微生物,楼主需要准备如下东东：
1、待测菌体.试管或培养皿都可以,但必须是纯粹的待测菌体,不能含有其他的微生物.
2、提纯的DNA.如果1有致病性或传染性,那么楼主就得自己提纯DNA再交给测序公司.DNA纯度要求可以做PCR.
我们在设计pcr引物的时候，经常需要将设计好的正反向引物与基因组DNA序列进行比对验证，但是因为反向引物是反向互补的，所以有些麻烦。因此，在这里介绍两个非常的小工具，一个是在线的模拟PCR，一个是本地运行的程序包，我会以附件形式上传，不需要蚁豆即可下载。两个都写得非常详细，相信大家都能看懂。不知道这个帖子能不能加分？希望版主看到后给予加分奖励
方法一：UCSC中的In-SilicoPCR
1、登录UCSC，网址链接http://www.genome.ucsc.edu/index.html，点击右侧工具的In-SilicoPCR，或者在Tools下拉框中点击In-SilicoPCR，如下所示：
2、点击In-SilicoPCR后，出现如下界面，我们需要将自己设计好的正向引物和反向引物分别粘贴至各自空白框，点击submit。对其它几个参数做点说明吧，MaxProductSize是指被放大区域的最大长度，MinPerfectMatch是指和引物的3‘末端PerfectMatch的最小碱基数目，MinGoodMatch:是指和引物的3‘末端GoodMatch的最小碱基数目，GoodMatch是3个碱基里最少匹配2个，FlipReversePrimer:反向引物反向互补。
3、我用自己设计的基因举例说明：HNF-1α的EXON2，参考序列：NM_000545，正向引物5’-3‘：CAGCCCTTGCTGAGCAGATCC，反向引物5’-3‘：CCACTGACTTCCTTTCCATCTACC。正向引物和反向引物分别粘贴至各自空白框，点击submit。出现如下结果，其中第一个红色标记指染色体位置，第二个红色标记指长度，第三个红色标记是我们输入的正反向引物，第四个标记可以直接链接至Primer3在线设计引物，网址http://primer3.ut.ee/。比对染色体位置就可以知道我们设计的引物是否包括HNF-1α的EXON2。
方法二：Reverse程序
这个程序是一位程序员朋友专门为我们编写的，操作非常简单，输入反向互补的引物序列后，就可以得到原始碱基序列。文件夹内一共才3个文件。
举例说明，HNF-1α的EXON2，参考序列：NM_000545，正向引物5’-3‘：CAGCCCTTGCTGAGCAGATCC，反向引物5’-3‘：CCACTGACTTCCTTTCCATCTACC。
1、点击target，粘贴反向引物序列。
2、点击右上角关闭键，这时候会弹出对话框，点击保存。
3、打开文件夹中的reverse_complement应用程度，按任意键即可自动关闭。
4、打开文件夹中的result，就可以得到反向互补引物序列对应的原始序列。
5、这时候就可以将正向序列和刚刚得到的原始序列一起代入HNF-1α的EXON2中检验我们设计的引物是否覆盖了目标片段。
好了，就这两个小工具了，非常实用，希望大家有所收获。有什么疑问可以给我留言或者私信我，支持我的战友就请给我投票吧。版主能给予我加分奖励吗？
reverse_complement.rar(95.82k)
非要说差别，可能就在测序引物是让片段线性增加，PCR引物是让片段指数增加。
需要在片段5‘端和3’端分别设计引物去扩增。
所以，对应的上游5‘端，我们习惯叫做”上游引物“，即forward primer，简称for；
对应序列的3’端，我们叫做下游引物，reverse primer，简称rev。
