This manual introduces the use of the SideStep II QRT-PCR Master Mix Kit, 1-Step product, including material preparation, operation process, precautions, etc.
The document details the use of the Agilent SideStep II QRT-PCR Master Mix Kit, which combines two modules to analyze gene expression in mammalian cells. The first module is a Cell Lysis Analysis Kit that allows for rapid preparation of lysates by skipping conventional nucleic acid purification steps, utilizing a buffer that stabilizes nucleic acids and inactivates cellular enzymes.
The second module is the Brilliant II QRT-PCR Master Mix Kit, 1-Step, used for probe-based detection protocols. The manual outlines specific procedures for preparing SideStep cell lysates, performing one-step QRT-PCR analysis on these lysates, normalizing variations in nucleic acid concentration using genomic DNA targets, and troubleshooting common issues.
| Reactions supported by normalization primers (Set 1) | fifty, 25-μl QPCR reactions |
|---|---|
| Reactions supported by QRT-PCR reagents | four hundred, 25-μl reactions |
| Storage temperature (upon receipt) | -20°C |
| Stability of SideStep lysates | at least 20 months when stored at –80°C |
| Guaranteed activity after thawing | 6 months |
They amplify distinct, single-copy regions of noncoding human genomic DNA to quantitate genomic DNA content for normalizing gene expression data or determining cell concentration.
They are suitable for analysis for at least 20 months when stored at –80°C.
No, it is for research use only and not for use in diagnostic procedures.