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ROX Reference Dye
ROX Reference Dye
ROX®为北京富百科生物技术有限公司注册商标，未经授权严禁使用!
ROX Reference Dye is used to normalize real-time RT-PCR and PCR assays. It provides an internal reference to which the reporter dye signal can be normalized. This signal normalization is necessary to correct for well-to-well signal fluctuations, often due to the design of the instrument.
Storage Condition : -20oC, protect from light
Shipping Condition : Ambient temperature
Product Description
ROX Reference Dye is particularly recommended for use in combination with FastStart TaqMan® Probe Master and FastStart SYBR Green Master on real-time PCR instruments from Applied Biosystems (ABI), in order to readily use their default parameter analysis. These instruments are available equipped with either a Halogen lamp-based light source (ABI PRISM 7000 Sequence Detection System, ABI 7300/7500 Real-Time PCR Systems) or a Laser-based light source (ABI PRISM 7700 Sequence Detection System, ABI 7900HT Real-Time PCR System). While both FastStart TaqMan Probe Master (Rox) and FastStart SYBR Green Master (Rox) can be readily used for the ABI 7500 System, it is necessary to increase the ROX concentration to 300 - 600 nM for the ABI 7000 System, the ABI 7300 System, the ABI 7700 System, and the ABI 7900HT System. ROX Reference Dye from Roche Applied Science is supplied in a “non-reactive” form, i.e., it is not able to react with e.g., amino groups of proteins like the ROX formulations supplied by other providers. In addition, it is proven not to inhibit PCR at a final concentration of up to 1000 nM. ROX Reference Dye is not required when using the LightCycler® Instruments.
Protocal
1. Add ROX Reference Dye to the FastStart Master (Rox) before primers, probes or DNA template are added. Depending on the volume of Fast- Start Master, add either undiluted ROX Reference Dye (1 mM) or a 1:10 diluted solution (0.1 mM) in order to avoid pipetting errors.
2. Adjust the final ROX concentration in the PCR mix to 300 - 600 nM. It is recommended to use initially a final concentration of
• 300 nM for the ABI 7000 System and the ABI 7300 System
• 400 nM for the ABI 7700 System and ABI 7900HT System
Ø In case you observe an unsteady fluorescence signal increase the ROX concentration until the signal is stable. Usually, when working with small reaction volumes (<10uL) a higher ROX concentration is required. (In this case, the recommended starting concentration is 400 nM.)
Ø Prepare the 0.1 mM ROX dilution by adding 450uL 0.1 M Tris, pH 8.0 to 50uL 1mM ROX.
Ø Although not recommended, you may also pipet the ROX Reference Dye directly to the PCR mix. In this case, prepare a 1:200 diluted (5 uM) ROX solution in 0.1 M Tris, pH 8.0. Add 2.5 or 3.4uL
Ø to 50 uL PCR mix containing FastStart TaqMan® Probe Master(Rox) or FastStart SYBR Green Master
Safety warnings and precautions
Warning: For research use only. These example protocols utilize chemicals that may be hazardous, and should only be performed by appropriately qualified and well-trained persons.
产品参数
| Name | ROX Reference Dye | ||
|---|---|---|---|
| CAT# | 023-50μL; 023-200μL | CAS# | N/A |
| Storage# | 4°C Sealed & desiccated & Minimized light exposure & Minimized Freezing And Thawing! | Shelf Life# | 12 months |
| Ex(nm)# | 568 | Em(nm)# | 591 |
| MW# | N/A | Solvent# | Water |
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