DyNAmo™ SYBR® Green qPCR Kit with ROX passive reference dye

DyNAmo™ SYBR® Green qPCR Kit with ROX passive reference dye

Description

DyNAmo SYBR Green qPCR kit with ROX passive reference dye (F-400RS, F-400RL) is a ready-to-use 2x master mix for sensitive, quantitative real-time PCR. DyNAmo 2x master mix contains a non-hot start DyNAmo DNA polymerase and all the other reagents needed for qPCR. Only template DNA and PCR primers need to be added by the user. ROX passive reference dye is also supplied with the kit in a separate tube for instruments that require ROX normalization.

The DyNAmo SYBR Green qPCR kit with ROX passive reference dye is compatible with all block-based real-time qPCR instruments. The DyNAmo kit has been developed in collaboration with MJ Bioworks, Inc

Advantages

	Sensitivity: Optimized Master Mix allows reproducible detection of low-copy number templates.
	Speed: More efficient amplification coupled with increased polymerase processivity results in earlier C(t)’s and shorter overall qPCR reaction times. (Figures 1 and 2) (Threshold cycle, Ct, the point at which the fluorescence signal is first recorded as statistically significant above background).
	Convenience: 2x master mix,one tube only. ROX passive reference dye provided in a separate vial.

DyNAmo kit - sensitivity, reproducibility and speed

Amplification curves below show clearly that lower copy number detection and earlier Ct-values are obtained with the DyNAmo kit. Signal level was also considerably higher and reaction times were substantially reduced when using the DyNAmo kit.

Figures 1a,2a: Amplification plot showing the linearity of the 10-fold dilutions of template as cycle numbers vs. fluorescence. Comparison of Finnzymes DyNAmo SYBR Green qPCR kit (1) and a qPCR kit from a leading supplier (2). A 221 bp PCR product was amplified from a plasmid containing a fragment of a human gene. Reactions were set up on the DNA Engine Opticon system (MJ Research, Inc) according to suppliers instructions. Reaction volumes of 20 µl were used with starting template copy number ranging from 1 to 1 000 000 copies.
Figures 1b,2b: Standard curves (right) generated from C(T) values. The curves can be used for measuring unknown amounts of sample target (the same target as used in standards).

Components

DyNAmo SYBR Green qPCR Kit with ROX passive reference dye	F-400RS	F-400RL
2x Master Mix (containing modified Thermus brockianus DNA Polymerase, SYBR Green I dye, optimized PCR buffer, 5 mM MgCl₂, dNTP mix including dUTP)	1 x 1 ml (for 40 reactions of 50 µl volume or 100 reactions of 20 µl volume)	5 x 1 ml (for 200 reactions of 50 µl volume or 500 reactions of 20 µl volume)
50x ROX passive reference dye	1 x 50 µl	1 x 250 µl
Instruction manual

ROX passive reference dye

ROX passive reference dye is used to normalize for non-PCR related fluorescence signal variation. Passive reference dye does not take part in the PCR reaction and its fluorescence remains constant during the PCR reaction. The ROX dye is provided as a 50x solution dissolved in a buffer that is compatible with the qPCR reaction buffer. Add the ROX passive reference dye to the qPCR master mix, 1 µl for each 50 µl reaction. At the 1x working concentration the ROX dye can be used to normalize data with ABI PRISM® real-time PCR instruments e.g. 7700 and 7900 SDS models. The amount of the ROX passive reference dye can be adjusted if different fluorescence level is needed with other instruments.

Storage

The DyNAmo qPCR kit is shipped in gel ice. Upon arrival, store all kit components at +4°C or -20°C. When stored and handled properly the DyNAmo kit is stable for six months in +4°C or one year in -20°C from the date of packaging.

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