DescriptionViPrimePLUS Lyophilized qPCR Master Mix is next generation first choice for a fast and easy real-time PCR reaction set up. The master mix is in freeze-dried form which produces stable preparation at room temperature. The freeze-dried master mix contains pure Taq DNA polymerases, MgCl2 and buffers components at optimal concentrations. ViPrimePLUS Lyophilized qPCR Master Mix has same optimal performance as ViPrimePLUS qPCR Master Mix and ViPrimePLUS qPCR Green Master Mix. The lyophilized qPCR master mix has highest sensitivity and specificity to detect the extremely low abundance transcripts with high efficiency. ViPrimePLUS Lyophilized qPCR Master Mix is used with supplied re-suspension buffer and ROX dye; hence hardware platforms that need ROX as a passive reference dye can be used in standard cycling conditions.
Features
- Convenient and rapid real time PCR reaction set up
- Lyophilized form – room temperature stable preparation
- Reliable quantification of low copy number targets
- High PCR sensitivity and specificity
- Compatible with most of the real-time PCR platforms
Kit Component
- 3 x Lyophilized qPCR MasterMix
- 1 x Lyophilized ROX
- 4 x Re-suspension buffer
Master Mixes Selection Guidelines for System CompatibilityTo calibrate a real-time PCR reaction, the various formulations of Master Mixes are available for most of the platforms.
| Real-time PCR platforms | ROX re-suspension volume | ROX addition per tube of master mix |
| Applied Biosystems 7000, 7700, 7900, 7300 StepOne, StepOnePLUS and ViA7 platforms, Roche capillary Lightcyclers. | 100µl | 20µl |
| All Stratagene platforms | 200µl | 15µl |
| Applied Biosystems 7500 platform | 700µl | 10µl |
| All other machines | Not Required | Not Required |
Ordering Information
| Catalog No | Description | Pack Size |
| QLMM05 | ViPrimePLUS Lyophilized qPCR Master Mix | 150 reactions |
DownloadManual
ViPrimePLUS Lyophilized qPCR Master Mix
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英 ['aisəutaip]
美 ['aisəutaip]
n. 同形像;
同形像统计图
选用和特异性抗体同种亚型的,同样荧光素标记的抗体,采用同样的染色步骤染同样浓度的细胞。然后上机检测。如果细胞有非特异性的结合,那同型对照的荧光强度可能会比未染色的阴性对照要高一些。
同型对照只用于那种阳性和阴性之间没有明确分解的情况。如果阳性细胞群非常清楚,是不需要同型对照的。另外,在多色同染的情况下,也不采用同型对照,而是采用FMO(fluorensence minus one)对照。
另外,对于同型对照的使用,也有很多反对意见。就算是同型抗体,也可能会有不同的亲和力,并不一定就能真实反应非特异性的结合。
同型对照(阴性对照)管:A同型+B同型+C同型
单阳性对照
A检测管:A+B同型+C同型
B检测管:B+A同型+C同型
C检测管:C+A同型+B同型
不存在荧光重叠的两个抗体不需要调节补偿。
样本为PBMC,想用三色标记Treg细胞,分别为CD4-PECY,CD25-Fitc,Foxp3-PE,想知道如何设置荧光补偿,用的是BD的流式仪,1、是设置阴性管一个,单阳性管每种一个,样品管一个么?
荧光素强度依次为PE, PECY5 APC FITC, PE最强,FITC最弱。 所以,建议抗体组合为:
Foxp3-PE,CD4-FITC,CD25-PECy5.5或者CD25-APC。
同时购买三个相应的同型对照抗体。
样品管设置如下:
1. cell + FITC-isotype + Pecy5.5( or APC)-isotype + PE-isotype
2. Cell + FITC-isotype + CD25-mAb + PE-isotype
3. Cell + PE-mAb + FITC-isotype + Pecy5.5( or APC)-isotype
4. Cell + CD4-FITC + Foxp3-PE + CD25 -PECy5.5( or APC)。 [这一管的数据可以拿来分析用]。
在充分了解调整电压和补偿原理的基础上,也可以做一些简化。如果不太懂,就按照这个来。其中Foxp3的抗体和同型对照抗体都是在固定破膜以后再加,不是与CD4或者CD25同时加。
英 ['aɪsətaɪp]美 ['aɪsəˌtaɪp]
n. 同型动物(或植物),图形文字,象征性图像;同位型;同模;同号模式
其同型为;percp-cy5.5(25ug包装)浓度:0.2mg/ml,
那么如果CD45RA抗体如果加入20ul,那同型抗体应该加入多少ml:
我自己的初步计算是:1ul的3分之1.不知道我算对了吗?应该怎么加才好呢?

