FlexStar® LDT (RT-)PCR Detection Mix 1.5
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Instructions for use FlexStar® LDT (RT-)PCR Detection Mix 1.5

FlexStar® LDT (RT-)PCR Detection Mix 1.5 For research use only! (RUO)

FlexStar® LDT (RT-)PCR Detection Mix 1.5

FlexStar® LDT (RT-)PCR Detection Mix 1.5 1. Application The FlexStar® LDT (RT-)PCR Detection Mix 1.5 is a buffered solution, which includes primers and a fluorescence labeled probe for the amplification and detection of the AltoStar® Internal Control 1.5. In combination with the FlexStar® (RT-)PCR Amplification Mix 1.5 the FlexStar® LDT (RT-)PCR Detection Mix 1.5 can be used for the real-time PCR based amplification and detection of nucleic acids. For research use only (RUO)! Not for use in diagnostic procedures. 2. Product content The FlexStar® LDT (RT-)PCR Detection Mix 1.5 contains the...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 • Repeated thawing and freezing of the No Template Control (NTC) (more than 4 times) should be avoided, as this might affect the performance of the product. • Storage at room temperature (max. +30 °C) should not exceed a period of 2 hours. • Protect the Detection Mix component from light. 4. Product description The FlexStar® LDT (RT-)PCR Detection Mix 1.5 is a reagent system. Used in combination with the FlexStar® (RT-)PCR Amplification Mix 1.5 and target specific primers and probes it allows the amplification and detection of nucleic acid. The...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 4.1 Components The FlexStar® LDT (RT-)PCR Detection Mix 1.5 contains enough reagents for 96 reactions. The product consists of the following components: • Detection Mix1) • NTC2) 1) Contains biological material of animal origin No Template Control (negative control) Except for the DNA polymerase and the reverse transcriptase, which are included in the FlexStar® (RT-)PCR Amplification Mix 1.5, the Detection Mix component contains all reagents (PCR buffer, magnesium salt, primers and probes) to allow detection of the IC. Primers and probes for the...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 5. Material required but not provided The following additional instruments and consumables are required for use of the FlexStar® LDT (RT-)PCR Detection Mix 1.5 but not provided with this product: • Appropriate real-time PCR instrument (see chapter 4.2 Real-time PCR instruments) • Appropriate nucleic acid extraction system or kit (see chapter 6.1 Sample preparation) • Vortex mixer • Centrifuge (e.g. desktop centrifuge) for centrifugation of kit reagents • Centrifuge for centrifugation of PCR plates • Appropriate 96 well reaction plates or reaction...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 For additional information and technical support regarding pre-treatment and sample preparation, contact altona Diagnostics technical support (see chapter 9. Technical assistance). 6.2 Preparation of the Detection Mix Add 30 µl specific primer/probe mix (not supplied by altona Diagnostics) per tube of Detection Mix. 6.3 Master mix setup All reagents and samples should be thawed completely, mixed (by pipetting or gentle vortexing) and centrifuged briefly before use. The FlexStar® LDT (RT-)PCR Detection Mix 1.5 is configured for use with the FlexStar®...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 ► Set up the master mix according to the following pipetting scheme: Table 2: Pipetting scheme (master mix setup) Number of reactions (rxns) Detection Mix (incl. specific primers/probes) NOTE No matter which method/system is used for nucleic acid extraction, never add the IC directly to the specimen. 6.4 Reaction setup ► Pipette 20 µl of the master mix into each required well of an appropriate optical 96-well reaction plate or an appropriate optical reaction tube. ► Add 10 µl of the sample (eluate from the nucleic acid extraction) or 10 µl of the...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 ► Thoroughly mix the samples and controls with the master mix by pipetting up and down. ► Close the 96-well reaction plate with appropriate lids or optical adhesive film and the reaction tubes with appropriate lids. ► Centrifuge the 96-well reaction plate in a centrifuge with a microtiter plate rotor and the reaction tubes in an appropriate centrifuge for 30 seconds at approximately 1,000 x g (~ 3,000 rpm). ► The NTC does already contain the IC template in the correct concentration. 7. Programming the real-time PCR instrument For basic information...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 7.2 Fluorescence detectors (dyes) ► Define the fluorescence detectors (dyes): Table 5: Fluorescence detectors Target Detector name Specific nucleic acid Depends on the specific primer/probe mix Internal Control 7.3 Temperature profile and dye acquisition ► Define the temperature profile and dye acquisition: Table 6: Temperature profile and dye acquisition Stage Cycle repeats Time [min:s] Reverse transcription 8. Data analysis For basic information regarding data analysis on specific real-time PCR instruments, refer to the user manual of the...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 8.1 Interpretation of results The Internal Control serves as (RT-)PCR inhibition control or as a control of the sample preparation procedure (nucleic acid extraction) and as a (RT-)PCR inhibition control. Detection of the IC in the JOE™ detection channel is not required for positive results in the specific detection channel. A high target specific RNA/DNA load in the sample can lead to reduced or absent IC signals. 9. Technical assistance For customer support, contact altona Diagnostics technical support: e-mail: 10. Trademarks and disclaimers...

FlexStar® LDT (RT-)PCR Detection Mix 1.5 11. Explanation of symbols Symbol Explanation Research use only Batch code Catalogue number Contains sufficient for "n" tests/reactions (rxns) Temperature limit Use-by date

FlexStar® LDT (RT-)PCR Detection Mix 1.5 Explanation Material number Version Note: Information is given to the user that is useful but not essential to the task at hand.