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Primer sets, length of amplicon, annealing temperature, extension time and PCR-protocol.

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https://figshare.com/articles/dataset/_Primer_sets_length_of_amplicon_annealing_temperature_extension_time_and_PCR_protocol_/192104
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PCR conditions Primer concentration: 2,5 µM of both forward and reverse primers. Buffer composition: LightCycler® 480 SYBR Green I Master. PCR volume 15 µl buffer and 5 µl DNA-eluate per PCR. PCR consumables White 96-well plate (Roche Diagnostics, Almere, The Netherlands) sealed with transparent self-adhesive foil (Roche Diagnostics, Almere, The Netherlands). PCR protocol Pre-incubation at 95°C for 10 minutes, de-annealing at 95°C for 10 seconds, 10 seconds annealing at the above-specified temperature, amplification at 72°C for the above specified time, 45 cycles. Melting-curve analysis: 5 seconds at 95°C, one minute at 65°C, 2.2°C per second increase of temperature until 97°C with five acquisitions per degree Celsius.
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2012-12-20
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