shelf life
≤6 mo.
packaging
kit of 1 mL (100 x 20 μL rxn; KK4670), kit of 10 mL (1000 x 20 μL rxn; KK4672), kit of 5 mL (500 x 20 μL rxn; KK4671)
manufacturer/tradename
Roche
compatibility
for use with Bio-Rad iCycler
shipped in
dry ice
storage temp.
−20°C
General description
KAPA SYBR® FAST One-Step qRT-PCR Kits are designed for optimal performance in stringent qRT-PCR reaction conditions, exhibiting dramatic improvements in sensitivity, specificity, reaction efficiency, and fluorescence. The kit is optimized for rapid one-step, one-tube RNA quantification, reducing experimental variation and contamination with a convenient qRT-PCR protocol.
KAPA SYBR FAST One-Step qRT-PCR Kits contain M-MuLV reverse transcriptase, RNase inhibitor, and a novel DNA polymerase evolved via directed evolution.
KAPA SYBR FAST One-Step qRT-PCR Kits contain M-MuLV reverse transcriptase, RNase inhibitor, and a novel DNA polymerase evolved via directed evolution.
Application
KAPA SYBR® FAST One-Step qRT-PCR Master Mix (2X) Bio-Rad® iCycler® is suitable for:
- Real-time polymerase chain reaction (RT-PCR)
- Gene expression analysis
- Low-copy gene detection
- Microarray validation
- miRNA research
- Gene knockdown validation
Biochem/physiol Actions
KAPA SYBR® FAST DNA Polymerase is inactive at ambient temperature. This prevents the formation of misprimed products and primer-dimers during reaction setup prior to the first denaturation step and results in high PCR specificity and accurate quantification. The RNase Inhibitor averts degradation of RNA target due to RNase contamination. Moloney murine leukemia virus reverse transcriptase (M-MuLV RT) has a high affinity for RNA and is optimized for cDNA synthesis at 42°C.
Features and Benefits
Increase signal and reaction efficiency:
Amplify across a broad range of target lengths:
Improve sensitivity and reproducibility:
Quick Notes:
- Consistently higher amplification efficiencies for accurate qPCR
- Higher fluorescence, earlier Cq values and improved reaction efficiencies
Amplify across a broad range of target lengths:
- Robust performance independent of amplicon size
Improve sensitivity and reproducibility:
- Accurate interrogation across a wide range of RNA template concentrations
Quick Notes:
- This kit contains wild-type M-MuLV and an engineered enzyme optimized for qPCR using SYBR Green I dye chemistry.
- The 2X master mix contains a proprietary buffer. Together with the novel enzyme, this improves amplification efficiency of both GC- and AT-rich targets.
- Use only gene-specific primers for one-step qRT-PCR.
- Optimal cDNA synthesis is achieved at 42°C for 5 min.
- 3 min at 95°C is sufficient for RT inactivation and DNA polymerase activation.
- For 3-step cycling, use 20 sec for primer annealing and 1 sec for extension/data acquisition at 72°C.
- Do not exceed 25 μL reaction volumes.
Preparation Note
Always ensure that components have been fully thawed and thoroughly mixed before use. The KAPA SYBR FAST qPCR Master Mix (2X) may not freeze solidly, even when stored at -20°C. The KAPA RT Mix is temperaturesensitive, and should be stored at -20°C and kept on ice during use.
The SYBR Green I dye contained in the KAPA SYBR FAST qPCR Master Mix (2X) and ROX/fluorescein dyes (depending on kit configuration) are light sensitive. Exposure to direct light for an extended period of time will result in loss of fluorescent signal intensity.
KAPA SYBR FAST qPCR Master Mix (2X) is stable through 30 freeze-thaw cycles. Ensure that all reagents are stored protected from light at -20°C when not in use. When protected from light, reagents are stable in the dark at 4°C for at least one week and may be stored at this temperature for short-term use provided that they do not become contaminated with microbes and/or nucleases.
The SYBR Green I dye contained in the KAPA SYBR FAST qPCR Master Mix (2X) and ROX/fluorescein dyes (depending on kit configuration) are light sensitive. Exposure to direct light for an extended period of time will result in loss of fluorescent signal intensity.
KAPA SYBR FAST qPCR Master Mix (2X) is stable through 30 freeze-thaw cycles. Ensure that all reagents are stored protected from light at -20°C when not in use. When protected from light, reagents are stable in the dark at 4°C for at least one week and may be stored at this temperature for short-term use provided that they do not become contaminated with microbes and/or nucleases.
Analysis Note
All kit components are subjected to stringent functional quality control, are free of detectable contaminating exo and endonuclease activity, and meet strict requirements with respect to DNA contamination.
Other Notes
For Research Use Only. Not for use in diagnostic procedures.
Legal Information
Bio-Rad is a registered trademark of Bio-Rad Laboratories, Inc.
SYBR is a registered trademark of Thermo Fisher Scientific or its subsidiaries
iCycler is a registered trademark of Bio-Rad
仅试剂盒组分
产品编号
说明
- qPCR Master Mix (2X) with fluorescein incorporated
- dUTP 10 mM
- 50X KAPA RT Mix
signalword
Warning
hcodes
Hazard Classifications
STOT SE 2
存储类别
12 - Non Combustible Liquids
wgk
WGK 1
flash_point_f
does not flash
flash_point_c
does not flash
法规信息
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An overview of directed evolution and the methods for generating proteins with optimized or entirely new functions.
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