ibGreen 100x for Real-Time PCR
ibGreen 100x for Real-Time PCR 100 µl
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ibGreen 100x for Real-Time PCR 100 ul
Cat.-No.: i05-G100 de 100 µl (100x)
ibGreen, an analog of SYBR® Green I, is a very sensitive dsDNA detection dye. High sensitivity, and high selectivi-ty for dsDNA allow to use ibGreen as a universal dsDNA detection reagent for qPCR. No need to use labeled probes to detect amplification with ibGreen - unlabeled primers are sufficient.
This formulation is specially designed to be used in real-time PCR experiments. Specific features are:
o Concentration of the dye is optimized for qPCR and carefully adjusted for reproducible results from lot to lot.
o PCR tested preparation
o Low fluorescence background - high fluorescence intensity gain
Storage conditions: Storage at -20°C in the dark. Transportation at room temperature for up to 3 weeks. Avoid prolonged exposure to light.
Protocol: qPCR with ibGreen
ibGreen, an analog of SYBR® Green I, is a very sensitive dye for the detection of double stranded DNA (dsDNA). Both dyes are used for non-specific detection of amplification in realtime qPCR experiments.
If the ibGreen reagent was stored below 0 ?, defrost it and keep it at room temperature. For rapid thawing, the reagent can be heated up to 50°?.
Calculate the volumes of reagents required for the reaction according to the manufacturer's instructions to PCR reagents. Please note that the ibGreen (100? in stock) must be diluted in a PCR master-mix of up to 1x concentration.
Prepare a 1x master mix containing no DNA according to the manufacturer's instructions to PCR reagents, add necessary amount of ibGreen. Keep the master mix and PCR tubes on the ice if you use Taq-polymerase without hot start.
Transfer the master mix to tubes or plates and add DNA.
Proceed with amplification according to your instrument manufacturer.
Always include positive and negative controls in your qPCR experiments. The temperature program for qPCR amplification does not differ from a standard PCR program for the given template and primers. For detection, the FAM or FAM/SYBR channel should be used. In order to be able to distinguish a specific product from primer dimers, use the melting curve step in your PCR program.