DyNAmo™ HS SYBR® Green qPCR Kit

DyNAmo™ HS SYBR® Green qPCR Kit

Description

DyNAmo HS SYBR Green qPCR Kit is a ready-to-use 2x master mix for highly specific quantitative PCR. The master mix contains a 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.

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

Advantages

	Specificity: DyNAmo hot start master mix minimizes the formation of unwanted primer-dimers and extension of non-specifically bound primers.
	Sensitivity: Optimized master mix allows reproducible detection of low-copy number templates.
	Broad dynamic range: DyNAmo HS detects as few as one copy of the target gene and achieves a dynamic range of up to eight orders of magnitude.
	Convenience: 2x master mix, one tube only. ROX passive reference dye provided in a separate vial.

DyNAmo HS kit - specificity and sensitivity

Amplification and melting curves below (Figures 1 and 2) show that DyNAmo HS kit has an excellent performance over a broad range of template copy numbers, and amplifies a specific product even from a difficult template.

Figures 1a and 1b. A 335 bp PCR product was amplified from Lambda DNA. Reactions were set up on the DNA Engine Opticon system (MJ Research, Inc). Reaction volumes of 20 µl were used with starting template copy number ranging from 5 to 50 000 000 copies.

Figures 2a-2c. A 481 bp amplicon of a beta actin gene was amplified with DyNAmo HS kit. Reactions were set up on the DNA Engine Opticon system (MJ Research, Inc). Reaction volumes of 20 µl were used with starting template amount ranging from 0.035 ng to200 ng. The melting curve shows that the DyNAmo HS kit has amplified the fragment with excellent specificity.

Components

DyNAmo HS SYBR Green qPCR Kit	F-410S	F-410L
2x Master Mix (containing modified Hot Start DyNAmo 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

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 HS SYBR Green qPCR kit is shipped in gel ice. Upon arrival store all kit components at -20 ºC. The DyNAmo HS SYBR Green Kit is stable for six months from the date of packaging when stored and handled properly.

After use, the leftover thawed 2x master mix can be refrozen and stored at -20 °C.

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