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• LightCycler^® 96 Instrument*
• LightCycler^® 480 Multiwell Plate 96, white (with sealing foils)*
• LightCycler^® 480 Sealing Foil*
• Standard swinging-bucket centrifuge containing a rotor for multiwell plates with suitable adaptors
• Nuclease-free, aerosol-resistant pipette tips
• Pipettes and multichannel pipettes with disposable, positive-displacement tips
• Autoclaved reaction tubes for preparing the master mix

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Vial / bottle	Label	Function / description	Content
1	LightCycler® PCR QC Kit, DNA Template, 15 ng/μl	Human genomic DNA	1 vial, 250 µl
2	LightCycler® PCR QC Kit, Primer Mix, 10x conc.	β-Globin primer pair mix	1 vial, 118 µl
3	LightCycler® PCR QC Kit, PCR Master, 2x conc.	FastStart Essential DNA Green Master	1 vial, 1,000 µl
4	LightCycler® PCR QC Kit, H2O, PCR Grade	To adjust final reaction volume.	1 vial, 1,000 µl

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Procedure	Time
PCR Setup	20 minutes thawing reagents and pipetting
LightCycler® 96 QC run	1 hour, 25 minutes
Total assay time	1 hour, 45 minutes

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• The SYBR Green I detection format is used to perform amplification and melting curve analysis. The test is comprised of 96 reactions to determine temperature accuracy across the entire block by calculating Tm values. Potential instrument effects on PCR results using Cq values are also measured. In addition, there is a check for fluorescence intensity by precisely monitoring the initial fluorescence between cycles 1 to 17.
• The LightCycler^® PCR QC Kit uses the FastStart Essential DNA Green Master, a ready-to-use reaction mix for hot start PCR in the SYBR Green I detection format. This master mix includes FastStart Taq Polymerase, a chemically modified thermostable recombinant Taq DNA Polymerase, showing no activity up to +75°C. This recombinant enzyme is active only at high temperatures at which primers can no longer bind nonspecifically. The enzyme is completely activated by removal of blocking groups in a single pre-incubation step (+95°C, 10 minutes) before PCR cycling begins. Activation does not require the extra handling steps typical of other hot start techniques.

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